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We narrowed to 325 results for: cloning

Showing: 161 - 180 of 325 results

  2. CRISPR-mediated Plant Base Editors

    Type
    Blog Post
    ... al., 2017). Our vectors have two Bsa1 sites to clone the single guide RNA (protospacer+scaffold) in a...single polycistronic tRNA-gRNA (PTG) gene system to clone multiple protospacers which enables us to target...

  3. Antibodies 101: Single Chain Fragment Variables (scFvs)

    Type
    Blog Post
    ...developed for another decade when in 1988 they were cloned by two separate labs (Bird et al., 1988; Huston...an scFv?  scFv sequences have traditionally been cloned by amplifying the VLand VH antibody sequences of...

  4. Five Popular Model Organisms, Part 2

    Type
    Blog Post
    ...around 20 minutes, they are perfect for molecular cloning and metabolic engineering.  The E. coli genome ...out the GreenGate Toolkit which allows for easy cloning of constructs specifically designed for use in ...

  8. CRISPR Plasmids - Empty gRNA Vectors

    Type
    Collection
    ... for Cloning 67720 Worm NotI/BamHI none S. pyogenes Colaiacovo pRB17 52527 Fly blunt end cloning none ...or nick. Selection , such as Puromycin or EGFP Cloning enzyme used for insertion of your gRNA sequence... by the Doudna Lab . Plasmid Expression System Cloning Enzyme Co-expressed Cas9 Cas9 System Selection ...BbsI yes, cut S. pyogenes Puro Zhang sgRNA(MS2) cloning backbone 61424 Mammalian BbsI none S. pyogenes ... U6 yes, cut (enhanced) S. pyogenes Zhang gRNA Cloning Vector Bbs I ver. 2 85586 Mammalian BbsI None S...nickase expression cassette using the Golden Gate cloning method. A separate accessory pack is available ...expressed from independent promoters. sgRNAs are cloned into separate vectors and then assembled into one...

  9. Open Enzyme Collection

    Type
    Collection
    ...Jennifer Molloy Lab Plasmids Blog: Golden Gate Cloning (Link opens in a new window) The Open Enzyme Collection...collection include polymerase chain reaction (PCR), cloning, isothermal amplification, reverse transcription...the basis of manufacturing their own enzymes by cloning them into an expression vector of their choice ...not functional by themselves. Researchers should clone these enzymes into an expression vector of their...

  11. Plasmid Cloning by PCR (with Protocols)

    Type
    Protocol
    ... Protocols Plasmid Cloning by PCR Plasmid Cloning by PCR You may also...Designing Primers for PCR Based Cloning The basic PCR primers for molecular cloning consist of: Leader Sequence... for primer design: Because we are cloning an ORF, we want to clone from the start codon (ATG) to the ...PCR based cloning carries a much higher risk for mutation than restriction enzyme based cloning. DNA replication...Protocols for plasmid cloning by PCR. Protocols...Ligation Bacterial Transformation Summary PCR based cloning is incredibly versatile and allows for nearly any...limitations. Background In its simplest form, PCR based cloning is about making a copy of a piece of DNA and at...

  12. Plan Your Experiment

    Type
    Collection
    ...using standard restriction-ligation cloning. However, the exact cloning strategy will depend on the gRNA ...including selecting the edit type, designing and cloning a gRNA, delivering Cas and the gRNA, and validating...Introduction Select Genetic Manipulation Design and Clone gRNA Select Expression and Delivery Validate Genetic...the multi-plasmid SAM system Activate Design and Clone Your gRNA Achieving your desired edit comes down...few factors to keep in mind when designing and cloning your gRNAs. Select Genetic Element to be Targeted... about how to design your gRNA . Synthesize and Clone Desired gRNAs Once your target sequences are selected...selected, it’s time to design your gRNA oligos and clone them into your desired vector. In many cases, you...

  14. CRISPR 101: Epigenetics and Editing the Epigenome

    Type
    Blog Post
    ...two vectors are part of the larger pMVP/pMAGIC cloning system, which contains Cas9 fusions for several...Arlotto ME, et al (2018) Creation of versatile cloning platforms for transgene expression and dCas9-based...

  15. CRISPR 101: Engineering the Plant Genome Using CRISPR/Cas9

    Type
    Blog Post
    ...researchers to insert targeting oligos with minimal cloning required. As with other model systems, multiple...containing vectors ready for Golden Gate mediated cloning of your gRNA of interest. Figure 1: Simplified...

  16. PiggyBac-ing Through the Genome Editing Field

    Type
    Blog Post
    ...cell lines is to transfect, select, and isolate clones. This procedure is cumbersome and inefficient, ... somewhere between one hundred and one thousand clones to find a cell line with the edit of interest. ...

  17. Ligation Independent Cloning

    Type
    Protocol
    ... Protocols Ligation Independent Cloning Ligation Independent Cloning You may also ...Independent Cloning (LIC) obviates the need for the time-consuming ligation step of traditional cloning methods... Independent Cloning Protocols...also like... Cloning Methods Empty Backbone Guide Molecular Biology Protocols Overview Ligation Independent...methods. In traditional cloning, base-pairing in the short overlapping regions (usually 4 bp) does not provide... restriction sites from the final product (no "cloning scars"). A “stuffer” sequence allows for electrophoretic... and may provide counter-selection for negative clones as in the example shown here. The manufacturer ...

  19. Protocols for Molecular Biology, Plasmid Cloning, and Viral Preps

    Type
    Protocol
    ...Video Restriction Cloning Assemble plasmids by restriction digest and ligation Cloning by PCR Generate ...single reaction Ligation Independent Cloning (LIC) Scarless cloning with Type II restriction enzymes and...and T4 polymerase pLKO.1 - TRC Cloning Vector Cloning protocols for using the pLKO.1 vector, a backbone...can be used for basic molecular biology, plasmid cloning, and titering and testing your viral preparations... the Lab Bench Basic Molecular Biology Plasmid Cloning Virus Antibodies Intro to the Lab Bench Introductory...tissues or cells without a commercial kit Plasmid Cloning Protocols for constructing and analyzing your plasmids...restriction sites by PCR Modification by Annealed Oligo Cloning Add a short stretch of DNA to a plasmid Gibson ...

  20. Protocol - How to Perform a Diagnostic Digest

    Type
    Protocol
    ...PCR - or restriction enzyme -based cloning to test individual clones before use of more expensive forms...digested with restriction enzymes common to multiple cloning sites, will result in similar sized bands, thus...Insert Orientation by Restriction Digest If you clone an insert into a vector by a single restriction ... enzyme, you will need to verify that it has be cloned in the correct orientation - this can be done by...restriction digest. Although this is never an ideal cloning strategy, sometimes it cannot be avoided. If you...therefore the only thing left to do is identify the clone(s) in which the insert is in the correct orientation... want to know how to differentiate between two clones that differ only in the orientation of the insert...
Showing: 161 - 180 of 325 results