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Overview of PCR Cloning
 
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PCR Cloning is an easy and reliable cloning method utilizing DNA amplification to generate the amplicon. Learn more at https://www.neb.com/applications/cloning-and-synthetic-biology/pcr-cloning
Views: 35247 New England Biolabs
PCR Cloning
 
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( http://www.abnova.com ) - PCR cloning is a method of cloning which dramatically reduces the time and effort put into the cloning reaction. PCR cloning procedure consisting of the four following steps: (1) production of a fragment of the gene using PCR, (2) digestion of genomic DNA, (3) ligation into a plasmid vector, and (4) transformation into bacteria and then bacteria will replicate the plasmid. More videos at Abnova http://www.abnova.com
Views: 45840 Abnova
Agarose Gel Electrophoresis
 
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For more information, visit http://www.bio-rad.com/yt/idea. This video demonstrates how to load and run DNA samples on an agarose gel. Basic information about the charge of DNA and how it will run in an horizontal electrophoresis cell is explained. http://www.bio-rad.com/evportal/destination/commerce/product_detail?catID=M5H6PI15&WT.mc_id=yt-lse-ww-biotech-20121012-vq759wKCCUQ We Are Bio-Rad Explorer. Our Mission: Bio-Rad’s Explorer program provides easy access to engaging hands-on science learning experiences that spark interest in science and its influence on the world. To do this we: Provide high quality, relevant, learning experiences based on real world science Empower educators with skills and confidence to deliver engaging, memorable lessons Connect with Bio-Rad Explorer Online: Website: http://www.bio-rad.com/en-us/education Twitter: https://twitter.com/BioRadEducation Pinterest: https://www.pinterest.com/bioradexplorer/ LinkedIn: https://www.linkedin.com/company/1613226/ Facebook: https://www.facebook.com/biorad/ Instagram: @BioRadLabs Snapchat: @BioRadLabs
Views: 416570 Bio-Rad Laboratories
Restriction enzymes
 
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Visit us (http://www.khanacademy.org/science/healthcare-and-medicine) for health and medicine content or (http://www.khanacademy.org/test-prep/mcat) for MCAT related content. These videos do not provide medical advice and are for informational purposes only. The videos are not intended to be a substitute for professional medical advice, diagnosis or treatment. Always seek the advice of a qualified health provider with any questions you may have regarding a medical condition. Never disregard professional medical advice or delay in seeking it because of something you have read or seen in any Khan Academy video.
Views: 230680 khanacademymedicine
Wizard® SV Gel and PCR Cleanup System - vacuum purification of PCR product
 
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http://www.promega.com/products/dna-and-rna-purification/dna-fragment-purification/wizard-sv-gel-and-pcr-clean_up-system/ : Quickly and easily purify DNA from PCR product using a vacuum manifold
Views: 3577 Promega Corporation
Simply Cloning - Supplement 1 - Troubleshooting
 
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In the case you don't get your clones from the first time, I want to suggest a couple of troubleshooting ideas. The first one deals with the case when you have no colonies on your vector plus insert plate on the next day after the transformation. In most cases the lack of colonies on the vector plus insert plate can be tracked to lost DNA, either of the vector or of the insert. To test that, I prepare an agarose gel and run 5 µl each of the vector and of the insert DNA. On this gel picture you can see relatively intense bands for both vector and insert. However, if one of your DNA fragments is barely visible or missing, it would most likely cause low efficiency of ligation. If that's the case, try to identify a step where you lose the DNA, or prepare the DNA fragment again. Another reason for not having any colonies on Day 2 is the low quality of plasmid DNA. An easy way to check the quality of plasmid DNA is to retransform 1 µl of the plasmid into competent cells. If you get a carpet of colonies like the one on this plate, then you plasmid DNA is good. However, if you get just 10 - 20 or no colonies, it would indicate the low quality of plasmid DNA. In such a case, pick one of those colonies, prepare fresh plasmid miniprep, and try to clone your insert into the fresh plasmid.
Views: 2973 Andriy Nemirov
Simply Cloning - Chapter 6 - Ligation
 
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Simply Cloning is a video manual for making DNA constructs. Chapter 6 describes how to ligate DNA fragments together using a T4 DNA ligase. Narration Script: During the ligation step we are going to mix the linerized vector and the insert together and add a T4 DNA ligase. The ligase will fuse their ends forming a circular plasmid. Ligation protocol: 1. Ligation mix o 9 µl ddH2O o 2 µl 10X ligation buffer o 2 µl of plasmid digested with restriction enzymes A and B o 6 µl of insert digested with restriction enzymes A and B. For the ligation control, use 6 µl of water instead of insert o 1 µl DNA ligase 2. Split ligation mix into two tubes 10 µl each 3. Incubate tube 1 at room temperature for 15 min 4. Incubate tube 2 at 16 oC for 24 hours Let's set up the ligation reaction. Here on the bench I have: • Double distilled water • A tube with cut vector • A tube with cut insert • An aliquot of ligase buffer - remember that the ligase buffer has to be aliquoted to avoid freeze-thawing • An empty tube for vector plus insert • An empty tube for control • And the T4 DNA Ligase I am going to set up two reactions, one with vector and insert and one with vector alone for control. In the Vector plus Insert tube I am going to combine: • 9 µl of water • 2 ul of cut vector • 6 ul of cut insert • 2 µl of ligase buffer • And 1 µl of DNA ligase And as always, I mix it up and down with a pipette. The control tube is pretty much identical, the only difference is that it contains water instead of insert. In the control tube I am going to put: • 15 µl of water, it's 9 + 6 instead of insert • 2 ul of cut vector • 2 µl of ligase buffer (we are skipping the insert) • 1 µl of DNA ligase And the mixing step. Now I am going to split the contents of ligation tubes into two aliquots. Here I have two new tubes, one of them I will label as Vector Plus Insert, another one as Vector Control. And I will transfer 10 ul from each of the ligation tubes into its replica. I am going to leave these two tube on the bench for 15 minute room temperature ligation, and I will put the other two tubes in a 16 oC incubator overnight. If tomorrow I will not get any colonies from the 15 minute ligation, I will repeat the transformation step with the tubes I kept at 16 oC.
Views: 13988 Andriy Nemirov
Simply Cloning - Chapter 1 - Planning
 
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Simply Cloning is a video manual for making DNA constructs. Chapter 1 deals with experiment planning, building plasmid maps in Vector NTI, and primer design Narration Script: In this section I am going to demonstrate how to create experiment outline in PowerPoint, build plasmid maps, and design primers for your cloning project. I begin each of my cloning projects by making a PowerPoint file with a project outline. For that I open a new PowerPoint file and save it as pSAT6-Bar, which is the project name. I change the layout to my favourite, which is "Title Only". And I put the project name on top. On this slide I put the name of my starting plasmid, which is pSAT6-MCS. Let's add some style to it. Then I draw a down arrow, add some style to it as well, and put the name of the plasmid I am trying to make, which is going to be pSAT6-Bar. For this one I will use a new style, which is going to be black. Also on this slide I want to have a name of my PCR template, which is a plasmid called pFGC 5941. Let's give it some style as well. Also I want to have the names of the primers I will be using, which is Bar-XhoI-F and Bar-BamHI-R. I will draw another arrow indicating that PCR product will be inserted into pSAT6-MCS. And, finally, I will indicate the restriction enzymes I will be using to cut my Vector and Insert. A usual cloning project goes for several days and in many cases you will be running several of those projects at a time. That's why I would recommend printing out this page and keeping it somewhere nearby, so when you come to your lab in the morning you know exactly where you left off. Let's move to the next part of the planning stage, which is building vector maps and designing primers. The software package I will use in this chapter is Vector NTI from Invitrogen. In Supplement 3, which is at the end of this video, I will demonstrate how to do exactly the same vector map design with a free software pDRAW32. I start with two browser windows, one displaying the sequence of pSAT6-MCS, another - sequence of pFGC5941. I launch Vector NTI Explorer, which keeps track of all the maps I have in the database. I create a new database subset, label it as Bar, and click on this icon, which says New. I name the molecule as pSAT6-MCS, indicate that it is circular, and open Edit Sequence window. Then I go back to the web browser, select and copy the sequence of pSAT6-MCS, and paste it in the Edit Sequence window. Let's look at this sequence in Vector NTI. Vector NTI opens with three panes. On the top right you see the map, on the bottom is the sequence, and on the top left is the information about the molecule. If you click on the Restriction/Methylation Folder you will see a list of all restriction enzymes currently displayed on the map. Now, let's add the features of pSAT6-MCS to this map. For that I click on ADD FEATURE, switch to the web browser window and select Feature View. You can see here a list of all pSAT6 features with their coordinates. Let's transfer them to our map. In the ADD FEATURE menu I am going to enter feature name, which is 2x35S Promoter, and starting and ending point. In the Vector NTI I could also pick different feature types, which reflect how the features are displayed on the map. What I particularly like about VNTI is that as I am adding those features, I can see them both on the map and on the sequence. Now I will proceed adding the rest of the features, namely Multiple Cloning Site, Terminator and Ampicillin resistance gene. Again, I get all the information about feature locations from the PubMed Nucleotide file. pSAT6-MCS map is ready, I am going to save it and move onto building pSAT6-Bar. I will build pSAT6-Bar by pasting the sequence of the bar gene from PubMed Nucleotide. First, I locate XhoI and BamHI sites on pSAT6-MCS and remove sequence between them. Then I will go to the PubMed Nucleotide and locate the sequence of the bar gene. When I scroll down and look in the annotations I can see that the bar gene is located on the reverse strand in the positions 373 - 924. To make life a little simpler, I will use PubMed tool called Change Region Shown. I enter here the boundaries of the bar gene and click on Update View. Now if I scroll all the way down you can see that the sequence of the pFGC5941 reduced to a 500 base pair region, which is the bar gene. Also, if you have a closer look at it, there are an ATG and a Stop codons on the complimentary strand of this sequence. So, let's copy the bar gene, go back to Vector NTI and paste this sequence between XhoI and BamHI sites in pSAT6-MCS.
Views: 49928 Andriy Nemirov
Designing PCR and Sanger Sequencing Primers– Seq It Out #5
 
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Download the free Sanger sequencing handbook at http://www.thermofisher.com/sangerhandbook Submit your question at http://www.thermofisher.com/ask Primer design is like art. There is more than one design to cover the region of interest. Are you an “Artist”? Primers are crucial to the success of target amplification and subsequent sequencing in PCR and Sanger sequencing workflows. Let’s take a look at our lab book In the typical Sanger sequencing workflow from genomic DNA, one needs to first amplify the target by PCR, and then subsequently run the Sanger sequencing reaction. If you start from purified plasmid DNA, one only needs to run the Sanger sequencing reaction. PCR amplification requires 2 primers from opposite strands that determine the region of sequence amplified in the forward and reverse direction. Sanger sequencing differs from PCR in that only a single primer is used in the reaction. Typically, for a given PCR fragment, two Sanger sequencing reactions are set up, one for sequencing the forward strand, the other one for sequencing the reverse strand. Primer design is an important aspect relating to many forms of PCR including basic PCR, fragment analysis, quantitative analysis and Sanger sequencing.. Here are a few things to keep in mind when designing your own primers. 1. Primer length should be in the range of 18 to 22 bases. 2. The primer should have GC content of 50% to 55%. 3. Primers should have a GC-lock on the 3’ end. 4. The melting temperature of any good primer should be in the range of 50OC to 55OC. 5. The primer should not include poly base regions. 6. Four or more bases that compliment either direction of the primer should be avoided. In addition, there are some PCR specific guidelines to help you design good PCR primers. These guidelines can be found on our website. Since primer design is not easy. There is risk of designing the wrong primers which could be costly in your experiments. You may be asking: Is there an easier way? The answer is Yes, and Thermo Fisher Scientific has a free tool to help you out. (www.lifetechnologies.com/primerdesigner) Primer Designer tool is a free PCR/Sanger primer online search tool that includes over 600,000 primer pairs covering the human exome and human mitochondrial genome. You can choose the range of amplicon length for your sample and your research interest to optimize it for your experiment. If you are doing NGS confirmation with your capillary electrophoresis genetic analyzer, this tool can really simplify your workflow. You can use this online by uploading your .vcf file, or if you have an Ion Torrent sequencer, even better, as there is a seamless integration with your ion reporter software to upload your data into this Primer Designer tool. Now you can find the right primers with just a few simple clicks. I hope this video was helpful on primer design, and I am sure you’ll have more questions. Submit your question at http://www.thermofisher.com/ask and subscribe to our channel to see more videos like this. And remember, when in doubt, just Seq It Out
Restriction Cloning
 
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SnapGene tutorial showing how to simulate standard cloning procedures.
Views: 38479 SnapGene
Key Steps of Molecular Cloning
 
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Sound track link http://incompetech.com/music/royalty-free/?keywords=tech+talk Script: Molecular cloning is a process of isolation of a specific DNA fragment and transfer of this fragment into a plasmid vector. As a part of the plasmid vector, the DNA fragment could be easily amplified, sequenced, stored for long periods of time, and used for gene expression and other functional studies. Two starting ingredients of molecular cloning are a plasmid vector and a DNA fragment that has to be inserted in it. The DNA fragment is usually a gene or other functional region from a living cell, or it could be an artificial sequence with properties useful for a researcher. Plasmid vector is circular piece of DNA that could be easily amplified in E. coli, stored for long periods of time, and easily manipulated in a test tube. A typical plasmid vector contains: - an origin of replication that allows it to be replicated inside a bacterial cell; - a selection marker, for example a beta lactamase gene coding for ampicillin resistance; - and a multiple cloning site , which could be cleaved with several restriction enzymes, such as BamHI (G-GATCC), EcoRI (G-AATTC) or NcoI (C-CATGG). In many vectors, the multiple cloning site is surrounded by sequences of promoter and terminator, that guide expression of inserted genes after the vector is introduced inside a cell. To be used for molecular cloning, both vector and insert DNA are treated with restriction enzymes that cleave double stranded DNA molecules producing overhanging single stranded nucleotide tails. After their ends have been prepared with restriction enzymes, vector and insert are combined together, and another enzyme, called a DNA ligase is added to the mix. At the same time as complimentary base pairing of single stranded overhands brings the ends of vector and insert together, the DNA ligase fuses them into one intact DNA molecule. In order to make multiple copies of this molecule, the ligation mixture is introduced inside the E. coli cells in a process called transformation. During the transformation the cell-DNA mixture is kept on ice and then exposed to 42 oC. Such sudden change in temperature drives the DNA inside some of the E. coli cells. Then the cells are plated on a plate with growth medium supplemented with a selective antibiotic. Only the cells that acquired the plasmid have resistance to the antibiotic and are capable of growth on such a medium. After overnight incubation at 37 oC each transformed cell produces a colony of identical cells, oftentimes called a clone. The selected clones are then individually picked, grown even further in a liquid medium, and the DNA is extracted from them. Thus, in the process of molecular cloning, a DNA fragment that represented a tiny fraction of cell genome is integrated into a bacterial plasmid. As a part of a plasmid, this DNA fragment represents a quarter or more of total DNA in a test tube and it could be effortlessly and endlessly amplified in E. coli.
Views: 233260 Andriy Nemirov
Blue white screening of DNA clones
 
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Lecture on blue white screening lacZ of DNA clones after cloning. http://shomusbiology.weebly.com/ Download the study materials here- http://shomusbiology.weebly.com/bio-materials.html The blue-white screen is a screening technique that allows for the rapid and convenient detection of recombinant bacteria in vector-based molecular cloning experiments. DNA of interest is ligated into a vector. The vector is then transformed into competent cell (bacteria), and the competent cells are grown in the presence of X-gal. Cells transformed with vectors containing recombinant DNA will produce white colonies; cells transformed with non-recombinant plasmids (i.e. only the vector) grow into blue colonies. β-galactosidase is a protein encoded by the lacZ gene of the lac operon, and it exists as a homotetramer in its active state. However, a mutant β-galactosidase derived from the M15 strain of E. coli has its N-terminal residues 11—41 deleted and this mutant, the ω-peptide, is unable to form a tetramer and is inactive. This mutant form of protein however may return fully to its active tetrameric state in the presence of an N-terminal fragment of the protein, the α-peptide. The rescue of function of the mutant β-galactosidase by the α-peptide is called α-complementation. In this method of screening, the host E. coli strain carries the lacZ deletion mutant (lacZΔM15} which contains the ω-peptide, while the plasmids used carry the lacZα sequence which encodes the first 59 residues of β-galactosidase, the α-peptide. Neither are functional by themselves. However, when the two peptides are expressed together, as when a plasmid containing the lacZα sequence is transformed into a lacZΔM15 cells, they form a functional β-galactosidase enzyme. The blue/white screening method works by disrupting this α-complementation process. The plasmid carries within the lacZα sequence an internal multiple cloning site (MCS). This MCS within the lacZα sequence can be cut by restriction enzymes so that the foreign DNA may be inserted within the lacZα gene, thereby disrupting the gene and thus production of α-peptide. Consequently, in cells containing the plasmid with an insert, no functional β-galactosidase may be formed. The presence of an active β-galactosidase can be detected by X-gal, a colourless analog of lactose that may be cleaved by β-galactosidase to form 5-bromo-4-chloro-indoxyl, which then spontaneously dimerizes and oxidizes to form a bright blue insoluble pigment 5,5'-dibromo-4,4'-dichloro-indigo. This results in a characteristic blue colour in cells containing a functional β-galactosidase. Blue colonies therefore show that they may contain a vector with an uninterrupted lacZα (therefore no insert), while white colonies, where X-gal is not hydrolyzed, indicate the presence of an insert in lacZα which disrupts the formation of an active β-galactosidase. Source of the article published in description is Wikipedia. I am sharing their material. Copyright by original content developers of Wikipedia. Link- http://en.wikipedia.org/wiki/Main_Page
Views: 90265 Shomu's Biology
Topo cloning
 
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Learn about topo cloning with this video. http://shomusbiology.weebly.com/ Download the study materials here- http://shomusbiology.weebly.com/bio-materials.html Source of all articles published in description is Wikipedia. Thanks to original content developers. Link- http://en.wikipedia.org/wiki/Main_Page TOPO cloning is a molecular biology technique in which DNA fragments amplified by Taq polymerase are cloned into specific vectors without the requirement for DNA ligases. The Taq polymerase has a nontemplate-dependent terminal transferase activity that adds a single deoxyadenosin (A) to the 3'-end of the PCR products. This characteristic is exploited in TOPO-cloning. The technique utilises the inherent biological activity of DNA topoisomerase I. The biological role of topoisomerase is to cleave and rejoin supercoiled DNA ends to facilitate replication. Vaccinia virus topoisomerase I specifically recognises DNA sequence 5´-(C/T)CCTT-3'. During replication, the enzyme digests DNA specifically at this sequence, unwinds the DNA and re-ligates it again at the 3' phosphate group of the thymidine base. TOPO vectors are designed in such a way that they carry this specific sequence 5´-(C/T)CCTT-3' at the two linear ends. The linear vector DNA already has the topoisomerase enzyme covalently attached to both of its strands' free 3' ends. This is then mixed with PCR products. When the free 5' ends of the PCR product strands attack the topoisomerase/3' end of each vector strand, the strands are covalently linked by the already bound topoisomerase. This reaction proceeds efficiently when this solution is incubated at room temperature with required salt.[1] Different types of vectors are used for cloning fragments amplified by either Taq or Pfu polymerase as Taq polymerase (unlike Pfu) leaves an extra "A" nucleotide at the 3'end during amplification. The TA TOPO cloning technique relies on the ability of adenine (A) and thymine (T) (complementary basepairs) on different DNA fragments to hybridize and, in the presence of ligase or topoisomerase, become ligated together. The insert is created by PCR using Taq DNA polymerase, a polymerase that lacks 3' to 5' proofreading activity and with a high probability adds a single, 3'-adenine overhang to each end of the PCR product. It is best if the PCR primers have guanines at the 5' end as this maximizes probability of Taq DNA polymerase adding the terminal adenosine overhang. Thermostable polymerases containing extensive 3´ to 5´ exonuclease activity should not be used as they do not leave the 3´ adenine-overhangs. The target vector is linearized and cut with a blunt-end restriction enzyme. This vector is then tailed with dideoxythymidine triphosphate (ddTTP) using terminal transferase. It is important to use ddTTP to ensure the addition of only one T residue. This tailing leaves the vector with a single 3'-overhanging thymine residue on each blunt end.
Views: 18283 Shomu's Biology
PCR - Polymerase Chain Reaction (IQOG-CSIC)
 
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PCR technique (Polymerase Chain Reaction), Animation. It is a technique used to make multiple copies of a segment DNA of interest, generating a large amount of copies from a small initial simple. Amplification of DNA segments makes possible the detection of pathogenic virus or bacteria, identification of individuals (DNA fingerprinting), and several scientific research involving DNA manipulation. Spanish version: http://youtu.be/TalHTjA5gKU This video has been produced in the Institute of General Organic Chemistry of the CSIC (IQOG-CSIC), Spain, by Guillermo Corrales, as part of its task for promoting Science Communication and may be freely used for educational and science popularization purposes. Canal Divulgación. Divulgación científica. Instituto de Química Orgánica General (QOG) CSIC Madrid, Spain http://www.youtube.com/user/CanalDivulgacion
Views: 1707937 CanalDivulgación
Transformation of E. coli with Plasmid DNA - Edvotek Video Tutorial
 
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For more information on transformation, check out our Quick Guide! http://www.edvotek.com/Transformation_Guide.pdf In the laboratory, scientists can force bacteria like E. coli to acquire exogenous DNA from the surrounding environment through a process called transformation. The newly acquired genetic information is both stable and heritable. Kits featured in this video: http://www.edvotek.com/223 http://www.edvotek.com/221 © 2013 Edvotek Inc. All rights reserved. www.edvotek.com
Views: 29848 Edvotek Inc.
Expression vector
 
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This lecture explains about the expression vector properties and the uses of expression vector in gene cloning. http://shomusbiology.weebly.com/ Download the study materials here- http://shomusbiology.weebly.com/bio-materials.html An expression vector, otherwise known as an expression construct, is usually a plasmid or virus designed for protein expression in cells. The vector is used to introduce a specific gene into a target cell, and can commandeer the cell's mechanism for protein synthesis to produce the protein encoded by the gene. The plasmid is engineered to contain regulatory sequences that act as enhancer and promoter regions and lead to efficient transcription of the gene carried on the expression vector.[1] The goal of a well-designed expression vector is the production of significant amount of stable messenger RNA, and therefore proteins. Expression vectors are basic tools for biotechnology and the production of proteins, for example insulin which is important for medical treatments of diabetes. An expression vector has features that any vector may have, such as an origin of replication, a selectable marker, and a suitable site for the insertion of a gene such as the multiple cloning site. The cloned gene may be transferred from a specialized cloning vectors to an expression vector, although it is possible to clone directly into an expression vector. The cloning process is normally performed in Escherichia coli, and vectors used for protein expression in organisms other than E.coli may have, in addition to a suitable origin of replication for its propagation in E. coli, elements that allow them to be maintained in another organism, and these vectors are called shuttle vector. Elements for expression Further information: Transcription (genetics) and Translation (biology) An expression vector must have elements necessary for protein expression. These may include a strong promoter, the correct translation initiation sequence such as a ribosomal binding site and start codon, a strong termination codon, and a transcription termination sequence. There are differences in the machinery for protein synthesis between prokaryotes and eukaryotes, therefore the expression vectors must have the elements for expression that is appropriate for the chosen host. For example, prokaryotes expression vectors would have a Shine-Dalgarno sequence at its translation initiation site for the binding of ribosomes, while eukaryotes expression vectors would contain the Kozak consensus sequence. The promoter initiates the transcription and is therefore the point of control for the expression of the cloned gene. The promoters used in expression vector are normally inducible, meaning that protein synthesis is only initiated when required by the introduction of an inducer such as IPTG. Protein expression however may also be constitutive (i.e. protein is constantly expressed) in some expression vectors. Low level of constitutive protein synthesis may occur even in expression vectors with tightly controlled promoters. Protein tags Main article: Protein tag After the expression of the gene product, it is usually necessary to purify the expressed protein. However, separating the protein of interest from the great majority of proteins of the host cell can be a protracted process. To make this purification process easier, a purification tag may be added to the cloned gene. This tag could be histidine (His) tag, other marker peptides, or a fusion partners such as glutathione S-transferase or maltose-binding protein. Some of these fusion partners may also help to increase the solubility of some expressed proteins. Other fusion proteins such as green fluorescent protein may act as a reporter gene. Source of the article published in description is Wikipedia. I am sharing their material. Copyright by original content developers of Wikipedia. Link- http://en.wikipedia.org/wiki/Main_Page
Views: 85231 Shomu's Biology
Troubleshooting a Bad PCR
 
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Synthetic Biology One is a free, open online course in synthetic biology beginning at the undergraduate level. We welcome scientists, artists, journalists, policymakers, or anyone interested in designing with DNA. Meet us at syntheticbiology1.com!
Simply Cloning - Supplement 3 - pDRAW32
 
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Illustration on how to build vector maps and design primers in pDRAW32 - free DNA editing program from AcaClone Software. Narration Script: pDRAW32 is a free DNA analysis software from the AcaClone Software. It does not have all the features of Vector NTI, but it is pretty good for simple DNA map building and primer design tasks. In this chapter I am going to use pDRAW32 to build maps of pSAT6-MCS and pSAT6-Bar and to design primers for amplifying the bar gene in PCR. To get started, I go to PubMed Nucleotide and find sequences of pFGC5941, which I will use as a PCR template for bar gene, and a map of pSAT6-MCS, which is the vector I will clone into. Then I launch pDRAW32, go to File - New - Enter New Sequence, and insert here the sequence of pSAT6-MCS. For that I go back to the web browser, scroll down, select the sequence, copy it, go back to pDRAW32 and paste it. Now I am going to add features to this sequence. I go to File - Edit and click on "DNA name, properties and annotations", where I can add the map features. I get the features of pSAT6-MCS from the PubMed file. I switch again to the web browser window, scroll up and select the Feature Table view. You can see a list of pSAT6 features with their coordinates. Let's transfer them to our map. For each feature I put its name, start and end position, orientation and type. Then I press on ADD. After all the features have been added I could check them with preview button. Then I switch to the DNA information tab. Here I insert the name of the plasmid and indicate that it is a circular DNA. Now it looks like a pretty map of pSAT6-MCS. Let's save it. Notice, that if you want to see the restriction sites on the plasmid map, you have to close and open pDRAW32. Now I am going to build a pSAT6-Bar map by pasting the sequence of the bar gene into the pSAT6-MCS. Let's first save this map as pSAT6-Bar. In pDRAW32 we need to separately edit the plasmid name, because it is not automatically imported from the file name. Now, let's go back to the PubMed Nucleotide and locate the sequence of the bar gene. When we scroll up and look at the annotations we could see that the sequence of thebar gene is located on the reverse strand in the positions 373 - 924. To make life a little simpler, I will use PubMed Option called Change Region Shown. I will insert here the coordinates of the bar gene and click on Update View. Now in my web browser I have the sequence of bar gene that I am going to copy. Before pasting this sequence into pDRAW32 I want to change its orientation by using a utility called Sequence Manipulation Tool. I paste here my sequence, select Reverse Compliment option and calculate the sequence. Here is the sequence of the bar gene, starting with the ATG and ending with the stop codon. I am going to copy it and hide this window for now. Editing a plasmid sequence in pDRAW32 is somewhat not trivial, but it can be done. First, I open a Sequence View window and locate the sites of XhoI and BamHI. Here we have CTCGAG, which is XhoI, and here we have GGATTC, which is BamHI. Please notice that those two restriction sites are located on lines beginning with nucleotides 1301 and 1351. Now I open an Edit Sequence window, locate XhoI site on the line 1301 and introduce a line break after it. Then I go to the line 1351, locate the BamHI site and introduce a line break before. Now I am going to remove the piece of Multiple Cloning Site between XhoI and BamHI and paste there bar sequence. Here on the map you can see a fragment that has been inserted between XhoI and BamHI sites. I am going to annotate this fragment as the bar gene. For that I will go to Edit - DNA annotations, and add a new feature, which would start in the position 1345 (right after the XhoI site) and would end in the position 1897 (right before the BamHI site). Now, let's design the PCR primers. With a plasmid map at hand it is quite easy. I am going to open a Sequence View window, hide all the ORFs, scroll down to the XhoI site, select 20 nucleotides right after it, and copy them with Control C. Then I open a Sequence Manipulation Tool and paste the sequence with Control V. I add six random nucleotides at the 5' end, select a restriction site, and hit Calculate. This is the primer sequence that I will send to an oligo synthesis company. Similarly, for the reverse primer, I will select the last 20 base pairs of bar, paste them into the Sequence Manipulation Tool, add a 5' overhang and a restriction site, check Inverted button and hit Calculate. Here I have shown you how to build plasmid maps and design PCR primers using some basic features of pDRAW32 .
Views: 17924 Andriy Nemirov
Eluator™ Vacuum Elution Device
 
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http://www.promega.com/products/dna-and-rna-purification/plasmid-purification/pureyield-plasmid-midiprep-system/ : The Eluator™ Device revolutionizes vacuum DNA elution using a midiprep or maxiprep.
Views: 697 Promega Corporation
Agarose Gel Electrophoresis - Amrita University
 
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▶ This video channel is developed by Amrita University's CREATE http://www.amrita.edu/create ▶ Subscribe @ https://www.youtube.com/user/amritacreate http://www.youtube.com/amritavlab ▶ Like us @ https://www.facebook.com/CREATEatAmrita ▶ For more Information @ http://vlab.amrita.edu/index.php?sub=3&brch=77&sim=1375&cnt=1 ▶ Amrita Virtual Lab Project website http://vlab.amrita.edu Agarose gel electrophoresis is the easiest and commonest way of separating and analyzing DNA. Nucleic acid molecules are separated by applying an electric field to move the negatively charged molecules through an agarose matrix. Shorter molecules move faster and migrate farther than longer ones because shorter molecules migrate more easily through the pores of the gel. This phenomenon is called sieving. The purpose of the gel might be to look at the DNA, to quantify it or to isolate a particular band. The DNA is visualized in the gel by addition of ethidium bromide. This binds strongly to DNA by intercalating between the bases and is fluorescent, meaning that it absorbs invisible UV light and transmits the energy as visible orange light.
Views: 220728 Amrita Vlab
Tutorial 2: Cloning Part 2 (Advanced PCR Primer Design)
 
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Constructing your expression vector with your DNA of interest. How to design primers for cloning. Leave your comments below and thanks for watching!
Views: 57533 ScytherScience
Causes of the smeared DNA bands after Gel electrophoresis and recommendations to avoid it
 
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Join Easy Experiment Academy for more lectures https://www.udemy.com/dna-primer-design-for-pcr/ Facebook page https://m.facebook.com/easyexperimentacademy/ -------------------------------------------------------- Gel electrophoresis, DNA Gel electrophoresis - Simple Animated Tutorial, Agarose Gel Electrophoresis, DNA gel electrophoresis lab demo, 2 D gel electrophoresis, Biotechnology - Gel Electrophoresis, Agarose Gel Electrophoresis of DNA fragments amplified using PCR, Principles of Gel Electrophoresis, PCR and Gel Electrophoresis , Performing Agarose Gel Electrophoresis, Gel Electrophoresis process, Gel Electrophoresis problems, Gel Electrophoresis procedure, Gel Electrophoresis principle, Gel Electrophoresis animation, Gel Electrophoresis explanation, Gel Electrophoresis steps, Gel Electrophoresis experiment, Gel Electrophoresis of dna, Gel Electrophoresis demonstration,
Views: 691 Shahed and Science
Primer Design for PCR
 
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In this lecture, I explain how to design working primers for use in PCR. If you are unfamiliar with PCR, watch the following video: http://www.youtube.com/watch?v=2KoLnIwoZKU. Created by:Tyler Maxfield
Views: 308520 Herbert Sauro
Competent Cell Transformation
 
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Visit http://www.invitrogen.com/compcells for more information. Overview of chemical transformation This video will walk you through the basics of chemical transformation. Transformation is the process by which bacterial cells take up foreign DNA from their environment. Typically, this is done in the lab for two main reasons: to propagate a recombinant plasmid or to obtain the results of a sub-cloning reaction. There are two main classes of competent cells: chemical and electro-competent. The procedure shown in this video can be used with most chemically competent cells. We are focusing on the protocol provided with Invitrogen's Top 10 strain of comp cell. Getting Started First, let's talk about storage conditions and what comes with your kit. Competent cells are stored at minus 80° degrees. Invitrogen's kits come with vials of competent cells, transformation instructions, a vial of S.O.C. medium, and a pUC19 transformation control. The OneShot® Top 10 kit used in this video comes with vials containing 50 micro liters of competent cells; enough for one transformation per tube. The competent cells must remain frozen until just before you are ready to use them. If they thaw too soon, it can affect the transformation efficiency. For your transformation protocol, you will need the following items: - A water bath set at 42° degrees - An ice bucket with ice - A 37° degree shaking incubator - A 37° degree incubator - 10 cm diameter LB Agar plates with the appropriate antibiotic In this case a 100 micrograms per microliter of ampicillin The selective plates should be warmed in an incubator for 30 minutes before you use them. You will need one plate for each transformation. Finally, you will need the vial of S.O.C. medium thawed and at room temperature. The first step is to mix the comp cells and the plasmid DNA of interest. Briefly centrifuge the DNA and put it on ice. This DNA can be from a ligation reaction or plasmid DNA you wish to propagate. Next, thaw on ice one 50 microliter OneShot™ vial of cells for each transformation reaction. Pipette one to five microliters of your DNA samples directly into each vial of competent cells. Mix by tapping gently and not by pipetting up and down. Your remaining ligation reaction can be stored at minus 20° degrees. We also highly recommend setting up a transformation control with the PUC 19 provided to ensure the transformation was performed correctly and the competent cells were at the expected competency. Add one microliter of the pUC19 control plasmid to 50 microliters of competent cells and tap gently to mix. The next step is to incubate the cells on ice for 30 minutes. Next is the heat shock step. This is harsh on the cells, so be sure not to mix or shake the vials. Incubate for exactly 30 seconds in the 42° degree water bath. After the incubation, remove and place on ice. Now the cells are allowed to recover from the transformation and are grown in rich S.O.C. medium. Add 250 microliters of the pre-warmed S.O.C. medium to each vial. Next, place the vials in a microcentrifuge rack inside the shaking incubator. Or, tape the vials on their sides to the shaking platform in the incubator. Shake the vials at 37°C for one hour at 225 rpm. If you do not have a shaking incubator you can add the vials to a 37°C regular incubator. Once the cells have recovered, it is time to plate them. The antibiotic you added will select only transformed cells. Pipette 15 microliters from the individual vial onto its own labeled LB plate. The remaining transformation mix may be stored at 4° and plated the next day if desired. For the control, remove 10 microliters from the vial and mix with 20 microliters of fresh S.O.C. and plate the entire 30 microliters as you did with the other transformations. To spread the cells evenly across the plate add 5 to 8 sterile glass beads and rotate, or use a sterile glass rod to spread across the plate. After removing the glass beads, invert the plates and incubate in 37° degrees overnight. Plasmid containing the antibiotic resistance marker that allowed them to grow in the presence of a selection antibiotic; the pUC19 control plate will have a larger number of colonies than your ligation plates. Many people also use blue white screening for selecting their clones. Each of these is a transformant and has taken up the plasmid containing the antibiotic resistance marker that allowed them to grow in the presence of the selection antibiotic. The pUC19 control plate will have a larger number of colonies than your ligation plates. Many people also use blue white screening for selecting their clones. For more trouble shooting tips or information on all competent cell structures visit http://www.invitrogen.com/compcells
Plasmid Mini Prep Protocol
 
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See more video protocols, please visit http://www.video-protocols.com MO BIO's UltraClean 6 Minute Mini and Standard Mini Plasmid Prep Kit - Detailed Protocol, including a description of "What's Happening" at critical steps.
Views: 9824 VideoProtocols Com
Plasmid as a cloning vector
 
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This dna cloning lecture explains use of plasmid as a cloning vector. http://shomusbiology.weebly.com/ Download the study materials here- http://shomusbiology.weebly.com/bio-materials.html A cloning vector is a small piece of DNA, taken from a virus, a plasmid, or the cell of a higher organism, that can be stably maintained in an organism, and into which a foreign DNA fragment can be inserted for cloning purposes.[1] The vector therefore contains features that allow for the convenient insertion or removal of DNA fragment in or out of the vector, for example by treating the vector and the foreign DNA with a restriction enzyme that creates the same overhang, then ligating the fragments together. After a DNA fragment has been cloned into a cloning vector, it may be further subcloned into another vector designed for more specific use. There are many types of cloning vectors, but the most commonly used ones are genetically engineered plasmids. Cloning is generally first performed using Escherichia coli, and cloning vectors in E. coli include plasmids, bacteriophages (such as phage λ), cosmids, and bacterial artificial chromosomes (BACs). Some DNA however cannot be stably maintained in E. coli, for example very large DNA fragment, and other organisms such as yeast may be used. Cloning vectors in yeast include yeast artificial chromosomes (YACs). Plasmid is an autonomously replicating circular extra-chromosomal DNA. They are the standard cloning vectors and the most commonly used. Most general plasmids may be used to clone DNA insert of up to 15 kb in size. One of the earliest commonly used cloning vectors is the pBR322 plasmid. Other cloning vectors include the pUC series of plasmids, and a large number of different cloning plasmid vectors are available. Many plasmids have high copy number, for example pUC19 which has a copy number of 500-700 copies per cell,[13] and high copy number is useful as it produces greater yield of recombinant plasmid for subsequent manipulation. However low-copy-number plasmids may be preferably used in certain circumstances, for example, when the protein from the cloned gene is toxic to the cells.[14] Some plasmids contain an M13 bacteriophage origin of replication and may be used to generate single-stranded DNA. These are called phagemid, and examples are the pBluescript series of cloning vectors. Source of the article published in description is Wikipedia. I am sharing their material. Copyright by original content developers of Wikipedia. Link- http://en.wikipedia.org/wiki/Main_Page
Views: 92777 Shomu's Biology
Creating and Analyzing Primers
 
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Need Benchling training documentation? Check out https://benchling.com/tutorials/25/creating-and-analyzing-primers. For DNA analysis tools, sign up at https://benchling.com
Views: 6252 Benchling
PCR  Vs Gene Cloning - Difference Between PCR And Gene Cloning
 
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What is the difference between a pcr primer and sequencing why do products have to be incorporated into plasmids simon' biology blog compare contrast pcr, vector cloning what answers how gene different from pcr? Understand bio 20 flashcards. What is the difference between pcr and cloning through plasmid polymerase chain reaction an alternative to molecular method dna in plasmids a technology for 21st centuryrecombinant genetic techniques university of leicester. Vector through a typical ligation reaction or by the action of an 'activated' vector that molecular cloning is set experimental methods in biology are used to because they contain foreign dna fragments, these transgenic similar polymerase chain (pcr) it fundamental difference between two pcr plasmids (jun 19 2011 ) soo, if principle os amplify gene interest (goi) and thereby produce many copies goi so when would you use one other feb 9, 2001 today, new forms related techniques about dozen different mutations per occur at fractions below decreasing variability experiments lab, such units early often taq. For example, if you clone the dna fragment in a sequencing vector, can use t7 or sp6 oct 16, 2014 sequence pcr products directly have single band but usually it is better to it, because of different reasons first your future plan gene interest into vector then 4, 2011 differences and similarities a)pcr vs cloning requires bacterial plasmid, does not. Is called polymerase chain reaction. Re when are pcr and bacterial cloning of dna used? . Oct 15, 2011 i need to understand the difference between polymerase chain reaction and plasmid vector cloning. The other type is in vitro which using the polymerase chain reaction (pcr) for vivo cloning a fragment of dna, containing single gene or number enables identification individuals and relationships between may 28, 2015 there any difference primer used pcr synthesized accourding to segment you want amplify. Vector through a typical ligation reaction or by the action of an 'activated' vector that in practice, process often involves combining dna different organisms. Difference between gene cloning and pcr a 'difference between' reference site. Pcr make the desired genes in cloning gene of interest along with molecular carrier is introduced into an expression system used that carries out dna replication over and again untill millions copies single or specifif sequence formed difference between pcr more perfect for teaching students to use technology a stem steam class biology forensics genomic library contains many different sequences; A clone primary polymerase chain reaction (pcr) traditional cdna double stranded using reverse transcriptase (or polymerase). Information about molecular cloning techniques, including restriction enzyme, two pieces of dna that have complementary overhangs, or which are both series plasmids allow you to do many different biology techniques in efficient hybridization between the 3' a overhang pcr product and. Pcr was invented by kary mullis in 1985gene cloning vs pcr sep 17, 2016. Difference between gene cloning and pcr difference youtubeplasmid vector wtf? ! help! biology reddit. Why use plasmid vector cloning in e coli to amplify a feb 6, 2001 fact, i've known several molecular biologists who didn't understand the differences and thought two techniques were interchangable, what is difference between pcr through vectors? In you take those copies of gene put them into that there was quick way doing it, cutting out middle man (or bug) an alternative cloning, called polymerase chain reaction(pcr), can be used at each round, number sequence primer sites for example, globin small sample dna isolated from screen genomic or cdna libraries clones encoding protein interest early often taq. I know they're both techniques used to amplify a specific dna sequence; Hell, i even carried out pcr in the lab this week.
Views: 829 MAD differences
pBR322 Gene Cloning Vector Explained
 
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Why the name pBR322? What are the elements in pBR322? Function of ori, restriction enzyme and selectable markers explained. Why pBR322 is known as 'ideal vector'? http://www.biologyexams4u.com/2013/01/gene-cloning-vector.html
Views: 81620 biologyexams4u
Design PCR primers to amplify a fragment that contains the mutations on msh2 in pmsh2 plasmid
 
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BIO125, Molecular Biology and Genomics, Spelman College, Spring 2014
Views: 685 Hong Qin
How to Prepare Your Samples for DNA Sequencing
 
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Erin Murphy, M.S., a Molecular Biologist with ACGT's DNA Sequencing group, demonstrates how to prepare your samples to ensure high quality DNA sequencing data.
Views: 11560 ACGT, Inc.
ApE PCR primer analysis, product size and mutation coverage
 
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Bio125, Molecular biology and genomics, Spelman College, spring 2015 Analyze PCR primers and verify their coverage of mutation on plasmid.
Views: 1159 Hong Qin
PCR Simulation
 
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Generate a PCR product using a pair of primers, detect non-specific amplification products, view PCR products on a gel and create a new project from the product.
Views: 990 Genome Compiler
Protocol 8: Miniprep (QIAprep Spin Kit)
 
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This tutorial explains how to carry out a miniprep using the QIAprep® Spin Miniprep Kit provided by Qiagen®. References: QIAprep Miniprep Handbook https://www.qiagen.com/resources/download.aspx?id=22df6325-9579-4aa0-819c-788f73d81a09&lang=en
Views: 18968 iGEM Academy
Overview of Traditional Cloning
 
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Traditional Cloning refers to the generation of DNA fragments using restriction enzymes, and their subsequent assembly and transformation. Learn more at https://www.neb.com/applications/cloning-and-synthetic-biology
Views: 8573 New England Biolabs
Gel Extraction Comb Trick
 
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In this video, Dan shows you a very simple and useful trick that you can use during your gel extraction/purification protocol. If you have lots of sample to load on the gel, and not a big enough well, you have to load into multiple wells. This means that you will have to cut out multiple bands, and still be careful to remove all the excess agarose. A better approach is to use a comb that will make wider wells. Don't have a big comb? Don't worry, Dan will show you how to make one using just your standard comb and some tape. Got your own trick? Leave a comment below! More videos: http://labtricks.com Got questions about lab work? http://www.labtricks.com/forum Other inquiries: [email protected] Find us on http://www.twitter.com/labtricks
Views: 11065 labtricks
Ligation
 
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THIS IS A GOOD VIDEO TO WATCH BEFORE DOING PCR. It mentions a lot of good tips for working with reagents and protocols. Specifically, this video goes over the Promega ligation protocol for Assignment 2. ***Note: In the video, Dr. Udall makes a control reaction following the "control" reaction mix on the protocol table. For the class, students will make a control reaction following the "background" reaction mix instructions. The background does not contain any DNA, just the vector. The ligation mix he makes is called "standard" in the protocol. This is a misnomer - be sure to refer to this experimental ligation mix by a more appropriate name.
Views: 2822 PWS310TA
Primer Design and Fragment Assembly Using NEBuilder HiFi DNA Assembly or Gibson Assembly
 
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Watch an interactive tutorial on primer design to see how simple it really is to clone with either NEBuilder® HiFi DNA Assembly or the Gibson Assembly® Cloning Kit.
Views: 20646 New England Biolabs
Expressioneering™ Technology: Surprisingly Simple Recombinant Protein Expression!
 
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Clone your favorite gene in an effortless afternoon, and start expression analysis the next day with the Expresso® Cloning and Protein Expression Systems, available with Rhamnose or T7 promoters and SUMO fusion tag to improve protein solubility. Expressioneering™ Technology uses in vivo homologous recombination to seamlessly clone PCR amplified DNA into specially designed expression vectors without the need for enzymes or purification steps. The desired insert is simply amplified with primers that include 18 bases that overlap with the ends of the Expresso vector. The unpurified PCR amplicon is then mixed with the pre-processed expression plasmid and the high-efficiency competent cells provided, and directly plated on appropriate media. Features of the Expressioneering Technology: • No vector preparation • No restriction enzymes or ligase required • No DNA purification steps • Tightly-controlled expression of N- or C-Terminal 6xHis-tagged proteins • Smallest vectors available for easier downstream manipulation (2.2 kb to 2.5 kb) See all of our Expresso products here: http://www.lucigen.com/Protein-Expression/
Views: 17891 LucigenVideo
Recombinant DNA technology | DNA Vectors | Cloning Vector And Expression Vector
 
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This recombinant DNA technology lecture explains about different types of DNA vectors such as cloning vector and expression vector. This lecture explains about the basic features of cloning vector and expression vectors and the difference between cloning and expression vector. In this video I also talk about different examples of cloning vector such as DNA plasmid, phagemid, cosmid, BAC, YAC and so on. For more information, log on to- http://www.shomusbiology.com/ Get Shomu's Biology DVD set here- http://www.shomusbiology.com/dvd-store/ Download the study materials here- http://shomusbiology.com/bio-materials.html
Views: 131106 Shomu's Biology
Gateway® Cloning Technology
 
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Visit http://www.lifetechnologies.com/gateway Gateway® recombination cloning technology circumvents traditional restriction enzyme based cloning limitations, enabling you to access virtually any expression system in just a few simple steps. From cloning or sub-cloning of DNA fragments over protein expression to functional analysis, Gateway® cloning technology is applicable for a variety of research areas, for truly multidisciplinary scientific studies and get's you there faster than with traditional methods. ------------------ Audio transcript: The typical cloning workflow involves many steps, takes a considerable amount of time and even then success is not guaranteed. Imagine a cloning workflow which was fast, flexible, efficient, and delivered consistent results. Well welcome to Gateway Cloning Technology. Regardless of the desired end result—cloning or subcloning of DNA fragments, protein expression, or gene analysis— Gateway Cloning Technology gets you there faster than traditional methods. Here's how it works. First, set up the reaction by adding the starting DNA material, buffer and clonase enzyme to your microfuge tube. Next, incubate at room temperature for one hour. Then add competent cells to your mixture and allow 45 minutes for the transformation to take place. After transformation, plate the reaction and incubate overnight at 37 degrees celcius. It is that simple! Gateway Cloning Technology gives you: • Fast, one hour, room temperature cloning reactions with greater than 99% efficiency. • Maintains orientation and reading frame without using restriction enzymes or ligation. • Eliminates re-sequencing ensuring consistent results throughout your experiment using the same clone from target identification to validation. • Shuttles DNA from one entry clone to multiple expression vectors, affording flexibility while simplifying your cloning workflow. Discover the versatility of Gateway Cloning Technology. Visit lifetechnology.com/gateway today.
How to transform E.coli via heat-shock method
 
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Transformation of competent E.coli cells via the heat-shock method. Reagents: LB Broth (1L): Tryptone - 10g Yeast Extract - 5g Sodium Chloride - 10g Mix, add DI H2O, autoclave, cool and use or keep in sealed autoclaved containers until needed. LB Agar (1L): Peptone -10g Yeast Extract - 5g Sodium Chloride - 10g Agar - 12g Dissolve in DI H2O, autoclave, wait until cool to the touch, add proper concentration of antibiotic if needed, pour 10-20ml per plate, allow agar to cool and harden. Use, or seal and keep at +4 until needed Protocol: 1. Thaw cells and DNA on ice (15-20min). 2. Mix DNA with cells, incubate on ice 15 min. 3. Heat shock at 42° C for 2 min. 4. Cool on ice for 3 min. 5. add 100ul LB, shake for 1 hour at 37° C. 6. Spread on plate 7. Place in incubator overnight at 37° C Any questions do not hesitate to ask. It is better to ask than waste reagents doing blind experiments
Views: 14704 bcburdoa
MIDSCI RapidTip PCR Purification in 60 Seconds (www.midsci.com)
 
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Visit www.midsci.com or call 800-227-9997 to order some today for your lab! RapidTip™ - PCR clean up in a tip in 60 seconds! Purification of PCR products for Sanger Sequencing in just one minute! No bind-wash-elute, enzymes, or magnetic beads necessary! An innovative new technology that removes undesirable impurities from PCR reactions leaving you with nothing but purified DNA for your Sanger sequencing applications. Primers and dNTPs will be gone in 60 seconds. Go to http://shop.midsci.com/scategory/M50/1772 for more details •Diffinity RapidTips - pre-packed with our proprietary material and ready to use out of the box! •A standard pipettor - single or multi-channel. •Microcentrifuge tubes to store purified DNA. •20-30µl PCR reaction volumes
Views: 192 MIDSCI Biotech
PCR Cloning - Simplify cloning in under 5 minutes!
 
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With In-Fusion® Cloning System efficiently construct any clone you desire, without subcloning. Learn more:http://goo.gl/5YkSS
Views: 7067 Takara Bio USA
Amplification and detection of foreign DNA (bacteria, GMO, etc.)
 
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Food & beverage labs often need to verify the absence of foreign DNA (bacteria, GMO, etc.) in food samples. In this workflow, the step of amplification and detection of the DNA is crucial. The power of PCR with its logarithmic amplification is still unbeaten for this task. Getting correct results quickly in this workflow means a lot in food production and delivery. For more about Food & beverage click here: www.eppendorf.com/food
Views: 2381 Eppendorf
Arcis DNA Isolation Video
 
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In less than 3 minutes with no prior sample preparation the Arcis DNA Prep range of products allow users to quickly and conveniently go from cellular material to downstream nucleic acid investigation without the need for traditional isolation or purification.
Views: 1533 Arcis Biotechnology