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We narrowed to 149 results for: pcr

Showing: 41 - 60 of 149 results

  1. Plasmids 101: In-house NGS Plasmid Quality Control Now at Addgene

    Type
    Blog Post
    Published
    April 6, 2021, 12:30 p.m.
    ...sample. The PCR is then performed, optimized as needed, and cleaned up using a standard PCR cleanup kit...scientists work with the depositing lab to develop a PCR strategy suitable for the library in question. We...that can amplify the insert of the library. This PCR step also adds the necessary adapter sequences for...intermediate steps, a sizing electrophoretic gel, and qPCR quantification of the final libraries. A broader...

  2. Plasmids 101: NGS Quality Control for Pooled Libraries

    Type
    Blog Post
    Published
    Oct. 26, 2017, 1:59 p.m.
    ...primers to create 200-300 bp sequencing products using PCR. A diagram of the product we created for the Brunello...library is shown in Figure 2. You can use a small PCR product because the plasmids in the library are identical...both pre- and post-amplification DNA and generates PCR sequencing products from both. These products are...

  3. Viral Vectors 101: Producing Your rAAV

    Type
    Blog Post
    Published
    July 16, 2024, 1:15 p.m.
    ...1999), quantitative PCR (qPCR) (Aurnhammer et al., 2012), droplet digital PCR (ddPCR) (Lock et al., 2014...Ehrhardt, A., & Baiker, A. (2012). Universal real-time PCR for the detection and quantification of adeno-associated...adeno-associated viral vector genome titers by droplet digital PCR. Human Gene Therapy Methods, 25(2), Article 2. https... experimental outcome and planning.  Both ddPCR- and qPCR-based titering assays measure physical titers...containing a viral genome. While ddPCR is superior in precision and variability, qPCR is the more widely used ...more details on the qPCR titering process, refer to the Addgene AAV Titration by qPCR Protocol. If you’re...interested in the ddPCR titering process, which is what Addgene uses, check out our ddPCR for AAV quantitation...

  5. Hassle-free 96-well Format Epitope Tagging Using Cas9 Ribonucleoprotein

    Type
    Blog Post
    Published
    June 28, 2018, 3:01 p.m.
    ...7 days. The crRNA and tracrRNA are annealed on a PCR block and the annealed cr:tracrRNA is then mixed ...using the csRNP method is as simple as doing routine PCR assays. Here is a simple to-do-list to follow when...Assemble csRNP: Anneal crRNA and tracrRNA using a PCR block. Add Cas9 protein to the annealed cr:tracrRNA...well plates: Characterize clonal lines by ICC and PCR genotyping (Sanger sequencing); use primers flanking... the tag sequence and expect a size-shift in the PCR product for knock-in lines. Troubleshooting low ...alternative tag (3xFLAG, Myc, V5, or HA). Tag-specific PCR genotyping can tell if HDR completely failed. If ...

  6. An “elegans” Approach to Better CRISPR/Cas9 Editing Efficiency

    Type
    Blog Post
    Published
    Dec. 17, 2020, 1:30 p.m.
    ...report that one can use linear repair templates (ie. PCR-derived dsDNA) with 30-60 basepair homology arms ...In the background are 504 restriction digests of PCR products used to identify a single 2xFLAG knock-in...used to generate linear dsDNA repair templates by PCR boosted efficiency (Ghanta et al., 2018). They further...screening for knock-ins much more efficient. Pure PCR repair templates are critical for efficient knock-ins...chromosomal gene modification with CRISPR/cas9 and PCR-based homologous recombination donors in cultured...

  7. Plasmids 101: Golden Gate Cloning

    Type
    Blog Post
    Published
    Aug. 27, 2015, 2:30 p.m.
    ...in the original plasmid (Option 1) or added using PCR-based amplification (Option 2). Advantages of Golden...work around this is to "domesticate" your fragment: PCR-based amplification can be used to create silent ...thus eliminating these from your gene of interest. PCR products are then digested with the Type IIS enzyme...

  8. Addgene's Top Blog Posts from 2020

    Type
    Blog Post
    Published
    Dec. 29, 2020, 2:15 p.m.
    ...Digital PCR for AAV Quantitation by Kate Harten DeMaio We made the switch to droplet digital PCR to quantify...

  9. Educational Planarian Tools for Teaching Developmental Biology Techniques

    Type
    Blog Post
    Published
    May 24, 2018, 1:20 p.m.
    ...create dsRNA. Students can produce DNA templates via PCR off of the same plasmid and use these to create riboprobes...can easily generate off of a circular plasmid via PCR. Typically, to make these constructs, developmental... create a complementary DNA (cDNA), and then use PCR and molecular cloning to insert the cDNA into the...

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

    Type
    Blog Post
    Published
    Oct. 8, 2020, 1:15 p.m.
    ...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...

  11. PITChing MMEJ as an Alternative Route for Gene Editing

    Type
    Blog Post
    Published
    Feb. 23, 2016, 3:30 p.m.
    ...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 ...

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

    Type
    Blog Post
    Published
    Sept. 28, 2017, 2:01 p.m.
    ...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...

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

    Type
    Blog Post
    Published
    Dec. 15, 2016, 3:57 p.m.
    ...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...

  15. RUBY-Red Siliques

    Type
    Blog Post
    Published
    March 1, 2022, 2:15 p.m.
    ...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...

  16. Intro to Our Summer SciComm Series!

    Type
    Blog Post
    Published
    June 9, 2022, 1:15 p.m.
    ...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...

  17. Tips for CRISPR Gene Editing in Mice

    Type
    Blog Post
    Published
    June 28, 2016, 10:59 a.m.
    ...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...

  18. 5 Great Apps for Lab Life

    Type
    Blog Post
    Published
    Sept. 22, 2016, 2:30 p.m.
    ...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...

  20. Antibodies 101: Conjugation

    Type
    Blog Post
    Published
    March 26, 2024, 1:15 p.m.
    ...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/...
Showing: 41 - 60 of 149 results