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We narrowed to 207 results for: pcr

Showing: 51 - 100 of 207 results

  1. Mycoplasma Contamination: Where Does It Come From and How to Prevent It

    Type
    Blog Post
    ...used for mycoplasma detection is using PCR. In this technique, PCR primers specific to the 16s RNA genes... you know it’s there? For this task, you can use PCR based detection, DNA staining, fluorescent tagging...Bae YJ, Hong JT, Chung YB, Lee CK, Song S (2006) PCR-based detection of mycoplasma species. J Microbiol...

  2. PITChing MMEJ as an Alternative Route for Gene Editing

    Type
    Blog Post
    ...homology required by MMEJ can easily be added via PCR amplification. Given the inefficiency of HDR for ...microhomologies are added to the GFP-Puro cassette via PCR, and this construct is inserted into the PITCh vector... 4: Select for puromycin resistant cells Step 5: PCR amplify and sequence the locus to verify correct ...

  3. Plan Your Experiment

    Type
    Collection
    ...cell population. The region of interest is PCR amplified, PCR products are denatured-renatured, treated... The PCR product is then run on an agarose gel to determine whether the edit was successful. PCR amplification...run on an agarose gel to identify DNA fragments. PCR and restriction digest (for HDR repaired DSBs): For...novel restriction site. The region of interest is PCR amplified, digested with the appropriate restriction...run on an agarose gel to identify DNA fragments. PCR amplification and gel electrophoresis (for HDR or...deletions or insertions, the region of interest can be PCR amplified using primers that (A) flank the region...and exact sequence of targeted alleles. Involves PCR amplification of targeted region from DNA, subcloning...

  4. AAVs CREATed for Gene Delivery to the CNS and PNS

    Type
    Blog Post
    ...inversion creates a template that’s suitable for PCR amplification using pre-designed primers. By using...interest. Repeat rounds of infection and screening by PCR enrich for capsid variants with increased ability...those that cross the barrier can be identified by PCR later. The PNS presents a different problem: it’s...

  6. Lambda Red: A Homologous Recombination-based Technique for Genetic Engineering

    Type
    Blog Post
    ...pairs.  dsDNA Substrate dsDNA inserts can be made by PCR using primers that amplify the DNA sequence of interest...recombineering substrates to avoid errors introduced during PCR. ssDNA Substrate ssDNA substrates can be generated...ordering synthetic oligonucleotides or denaturing short PCR products. Either way, the substrate should be ~70...the presence of the desired modification. Colony PCR can be used to screen for positive clones in most...

  7. RUBY-Red Siliques

    Type
    Blog Post
    ...selection would only then kill the desired plants! PCR or fluorescent markers are currently used in these... construct could be achieved much easier than by PCR or a fluorescent microscope. First, red CRISPR-positive...

  8. Intro to Our Summer SciComm Series!

    Type
    Blog Post
    ...definition) in a specific field.  For example, while “PCR test” has recently transitioned from jargon to non-jargon...jargon in most other uses. Many laypeople would use “PCR test” to mean “a test that uses a biological sample...

  9. Tips for CRISPR Gene Editing in Mice

    Type
    Blog Post
    ...primer sets to use to genotype your pups. A simple PCR with these primers will let you know if there are...are any major deletions, and sequencing of the PCR products will give specific details on what exactly ...alleles are present in these mice, you can clone the PCR products into a pBluescript cloning vector and sequence...

  10. 5 Great Apps for Lab Life

    Type
    Blog Post
    ...use protocols that require restriction enzymes or PCR. The app contains a double digest tool that is only...Lastly, the Tm calculator helps you set up an optimal PCR depending on the primer sequence and the NEB polymerase...

  12. Viral Production

    Type
    Collection
    ... AAV particles are titered by droplet digital PCR (ddPCR) using primers and probes targeting the ITR elements...April 2016) also performed by droplet digital PCR (ddPCR). Quality Control Addgene ensures high quality...are titered using a lentiviral droplet digital PCR (ddPCR) titration protocol (modified from Wang et al...confirmed to have the correct integrated provirus using PCR. This assay involves transducing cells with serial...obtained using ddPCR are generally 2–3 fold higher than those achieved using the standard qPCR method. Titering...

  13. Antibodies 101: Conjugation

    Type
    Blog Post
    ...conjugates used in assays like proximity ligation, immuno-PCR, and single-cell applications (Hegazy, et al., 2020...(2007) Detecting antigens by quantitative immuno-PCR. Nat Protoc 2:1918–1930. https://doi.org/10.1038/...

  14. Transferable Skills Guide: Problem Solving

    Type
    Blog Post
    ...plagued by a steady barrage of problems. Why didn’t my PCR work? Why didn’t my ligation work? Why are there ... being able to figure out what went wrong with a PCR is a pretty niche skill; outside of a lab, where ...

  15. Summer SciComm Series: Cognitive Load

    Type
    Blog Post
    ... once decided that to teach high school students PCR, they first had to understand hydrogen bonds and ...unlikely to be able to follow a YouTube video on PCR, while a senior lab scientist is likely to be quickly...

  17. Zhang Lab's CRISPR Frequently Asked Questions

    Type
    Collection
    ...better for PCR and surveyor. I used DNA polymerase Takara Ex Taq ™ DNA Polymerase for my genomic PCR, but couldn't...conditions with 5% DMSO or without 5% DMSO. My genomic PCR didn’t work, do you have any suggestions? The Zhang...the protocol, then uses around 2-4ul in each 50ul PCR reaction. The gDNA extracted in this way is usually...groups have successfully used Herc II or Kapa Hifi in PCR. Maybe this Takara enzyme is not very robust in this...

  19. Tips for arabidopsis transformation

    Type
    Blog Post
    ... Weigel and Glazebrook, 2002. Tips: Do a colony PCR on your agrobacterium: Getting a transformant takes...remaining starter culture can be used for colony PCR and glycerol stocks. Dip/drip plants more than once...

  22. Getting the Most from Your Lentiviral Transduction

    Type
    Blog Post
    ... use detector cell lines, ELISA, fluorometry, or PCR-based approaches to detect even low levels of contamination...detection, Uphoff et. al. outline a do it yourself PCR-based approach complete with freely available positive...

  24. Plasmids 101: Biotinylation

    Type
    Blog Post
    ...Reaction (PCR) by using a biotinylated primer. Biotin can also be enzymatically incorporated during PCR using...

  25. PiggyBac-ing Through the Genome Editing Field

    Type
    Blog Post
    ...differentiation process. Moreover, a splinkerette-PCR assay may be used to map the specific genetic loci...and D. J. Adams, “A high-throughput splinkerette-PCR method for the isolation and sequencing of retroviral...

  28. Plasmids 101: What is a plasmid?

    Type
    Blog Post
    ...single-stranded DNA sequence used as an initiation point for PCR amplification or sequencing. Primers can be exploited...

  30. 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 ...

  31. CRISPR Plasmids - Tagging

    Type
    Collection
    ... technique for use in Drosophila cells that uses PCR to generate both an expression cassette for the Cas9.... The plasmids in the following articles provide PCR templates for amplification of the tag (eg GFP, Flag...expresses Cas9, PITCh-gRNA, & FBL-specific gRNA pCRIS-PITChv2-FBL - PITCh donor vector for EGFP-2A-PuroR...

  32. 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...

  36. 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...

  37. 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...

  38. Plasmid Cloning by PCR (with Protocols)

    Type
    Protocol
    ...3’. Experimental Procedure Run PCR and Ppurify 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...

  39. Targeted Mutagenesis with EvolvR

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

  40. 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 ...

  42. CRISPR Plasmids - C. elegans

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

  44. 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...

  45. 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...

  48. Working with Nuclear Receptors

    Type
    Blog Post
    ...nuclear receptor and coregulator research. Our Q-PCR datasets, for example, document the expression patterns...

  49. 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...
Showing: 51 - 100 of 207 results