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Enter a sequence to perform a BLAST-based search of Addgene plasmid sequence databases.

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We narrowed to 210 results for: pcr

Showing: 81 - 100 of 210 results

  1. Zhang Lab CRISPR Page

    Type
    Collection
    ... be co-transfected as PCR amplicons using a protocol similar to SpCas9 sgRNA PCR (Ran et al., Nature Protocols...codon-optimized SpCas9; sgRNA can be co-transfected as PCR amplicons (Ran et al., Nature Protocols 2013). Individual...

  2. Selecting Your Plasmid Purification Kit

    Type
    Blog Post
    ...DNA quality can inhibit important reactions like PCR or restriction digests. Transfection Grade DNA (high...from tissues, blood, virus, or other samples, for PCR and gel cleanup, and for extracting RNA! (That’s ...

  3. What is Polymerase Chain Reaction (PCR)

    Type
    Protocol
    ... Reaction (PCR) Polymerase Chain Reaction (PCR) You may also like... Plasmid Cloning by PCR Restriction... how to perform Polymerase Chain Reaction (PCR). Basic PCR Program Initial Denaturation for 2 minutes ...strands. Materials List Reagents for each 50µL PCR: Thin-walled PCR tube Ice Bucket 2 μL Template DNA (10 ng...Polymerase (5 units/μL) PCR Machine Agarose Gel Procedure Primer Design and PCR Design Primers. See our...volume of master mix to add to each PCR tube. Secure the tops to the PCR tubes, gently tap each tube to bring...cloning by PCR to learn how to design primers for cloning purposes. What do I do if my PCR isn't working...Information A standard Polymerase Chain Reaction (PCR) is an in vitro method that allows a single, short...

  4. AAV ddPCR Titration

    Type
    Protocol
    ... Bio-Rad, 1863051 8-strip PCR tubes, Axygen, PCR-02-FCP-C ddPCR 96-well PCR plates, Bio-Rad, 12001925 ...302411100 Droplet Digital PCR System, Bio-Rad, DX200 Thermal Cycler, Bio-Rad, T100 PCR Plate Sealer, Bio-Rad...20X): 5 µL in 95 µL 1X PCR buffer (1:20) Dilution 2 (20X): 5 µL in 95 µL 1X PCR buffer (1:400) Dilution...20X): 5 µL in 95 µL 1X PCR buffer (1:8,000) Dilution 4 (20X): 5 µL in 95 µL 1X PCR buffer (1:160,000) Dilution...: 5 µL in 95 µL 1X PCR buffer (1:3,200,000) Dilution 6 (2X): 50 µL in 50 µL 1X PCR buffer (1:6,400,000... 50 µL in 50 µL 1X PCR buffer (1:12,800,000) Dilution 8 (2X): 50 µL in 50 µL 1X PCR buffer (1:25,600,000...well PCR tube strip into a chilled 96-well freezer block. Add 20 µL of the master mix to each PCR tube...

  5. Plasmid Cloning by PCR (with Protocols)

    Type
    Protocol
    ...-3’. Experimental Procedure Run PCR and Purify the PCR Product Run PCR to amplify your insert DNA. It ... cDNA library, or by RT-PCR. Isolate your PCR product from the rest of the PCR reaction using a kit, such... Protocols Plasmid Cloning by PCR Plasmid Cloning by PCR You may also like......recipient plasmid. Designing Primers for PCR Based Cloning The basic PCR primers for molecular cloning consist...Link opens in a new window) QIAquick PCR Purification Kit . The PCR product is now ready for restriction... your PCR product is the anticipated size and that your band is strong (indicating that the PCR reaction...for plasmid cloning by PCR. Protocols...

  9. CRISPR Plasmids - C. elegans

    Type
    Collection
    ...need Cas9 plasmid Joung 46170 PU6::klp-12_sgRNA cU6 PCR (see paper) Transfection none, need Cas9 plasmid ...

  11. CRISPR Plasmids - Empty gRNA Vectors

    Type
    Collection
    ... none S. pyogenes Kamoun pUC119-gRNA 52255 Plant PCR template none S. pyogenes Sheen pRGEB31 51295 Plant...none S. pyogenes Boxem PU6::klp-12_sgRNA 46170 Worm PCR (see paper) none S. pyogenes Calarco SP6-sgRNA-scaffold...pyogenes Sternberg pJW1311 61254 Worm none, fusion PCR none S. pyogenes Ward pCFD1-dU6:1gRNA 49408 Fly BbsI...Genome Editing in Monocot and Dicot Plants Plant PCR-based procedure to rapidly generate multiple sgRNA... pyogenes Conklin pcrRNA.ind 61284 Bacteria araB none S. pyogenes Beisel pcrRNA.con 61285 Bacteria J23119...Enzyme Co-expressed Cas9 Cas9 System Selection PI pCRISPR 42875 Bacteria BsaI none S. pyogenes Chloramphenicol...42250 Zebrafish BsaI none S. pyogenes Joung pCRISPathBrick 65006 Bacteria BsaI yes, interfere S. pyogenes...

  12. Targeted Mutagenesis with EvolvR

    Type
    Blog Post
    ...(nCas9). Similar to mutagenesis with error prone PCR, EvolvR uses an error prone polymerase to introduce...

  13. Antibodies 101: ChIP

    Type
    Blog Post
    ...the crosslinking. Finally, purify your DNA with a PCR purification kit or with a phenol-chloroform extraction...through a variety of downstream methods, such as qPCR or next-gen sequencing. ChIP is an incredibly useful...isolated and purified DNA, such as sequencing or qPCR. Yay!  Where’s the challenge in that?  So while ...

  15. Gibson Assembly Protocol

    Type
    Protocol
    ...as point mutations). Generate DNA segments by PCR. Run PCR product on an agarose gel to check for size ...-purify DNA segments. Otherwise, PCR purification or even the raw PCR mix can work fine in an assembly...figures). These identical sequences can be created via PCR with primers that contain a 5′ end that is identical...This trick can also enable replacement of "inverse PCR" reactions with a two-part Gibson reaction if you're...oligos When you need intervening sequence between two PCR products, one method is to “stitch” together several...be inserted is too long to include on overlapping PCR primers (>60 bp) but too short to make its own part...the Gibson reaction are different than with normal PCR primers (Gibson et al. , 2010). Number of fragments...

  16. Lentivirus ddPCR Titration

    Type
    Protocol
    ... Bio-Rad, 1863051 8-strip PCR tubes, Axygen, PCR-02-FCP-C ddPCR 96-well PCR plates, Bio-Rad, 12001925 ...5425R Droplet digital PCR System, Bio-Rad, DX200 Thermal Cycler, Bio-Rad, T100 PCR Plate Sealer, Bio-Rad...well PCR tube strip into a chilled 96-well freezer block. Add 16 µL of the master mix to each PCR tube...perform digital droplet PCR (ddPCR) for lentivirus vector titration. Protocols...protocol describes how to use droplet digital PCR (ddPCR) to titer lentivirus vectors. This protocol specifically... 4 µL of the 25 ng/µL samples to the appropriate PCR tubes. Pipette back and forth 10 times to mix. Generating... droplets. Transfer the droplets to a prechilled PCR plate. Pro-Tip To ensure that the droplets are not...

  17. Plan Your Experiment

    Type
    Collection
    ...methods like sequencing or digital PCR , RNA-based methods like qPCR , and/or protein-based methods like...Very inefficient Can't distinguish specific edits PCR Knockouts Large insertions/deletions Some prime and...provide specific sequence information alone Digital PCR Knockouts Insertions/deletions Prime and base edits...RNA-sequencing, Perturb-seq, deep sequencing, etc. qPCR Knockouts Activation/interference Needs good primer...
Showing: 81 - 100 of 210 results