Skip to main content
Addgene

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

In Category

We narrowed to 963 results for: tes

Showing: 461 - 480 of 963 results

  1. Antibodies 101: Immunoprecipitation

    Type
    Blog Post
    ...the immunoglobulin, leaving the antigen binding sites free to capture the target protein. The beads can...indiscriminately, causing competition for binding sites. In cases like these, antibodies can be directly...sample. It is important to remember that sample lysates will also contain proteases and phosphatases that...sample.  Eluting the target protein for downstream testing Once the non-specific proteins are removed through...

  2. CRISPR 101: Targeting RNA with Cas13a (C2c2)

    Type
    Blog Post
    ... Cas13a binds and cleaves RNA rather than DNA substrates. In terms of structure, Cas13a shares no homology...molecule through a uracil-rich stem loop and facilitates target binding and cleavage through a series ...changes in the Cas13a molecule. Like Cas9, Cas13a tolerates single mismatches between the crRNA and target...PMC4660269. 7. Joung J, Ladha A, et al., ‘Point-of-care testing for COVID-19 using SHERLOCK diagnostics.’ medRxiv...

  3. Getting the Most from Your Lentiviral Transduction

    Type
    Blog Post
    ...may include reduced saturation density, decreased rates of growth, or clumping. To ensure the best possible... culture-based experiment, labs should routinely test for mycoplasma contamination. There are several ...with freely available positive controls. Routinely test your cell line for mycoplasma and discard it if ...routinely produce lentivirus may want to consider testing a variety of cation and serum sources to ensure...

  4. Plasmids 101: E. coli Strains for Protein Expression

    Type
    Blog Post
    ... (pLemo) Rhamnose-tunable T7 RNAP expression alleviates inclusion body formation. Expression vector cannot... recA+, KmR Expression of toxic proteins  * Denotes the presence of an additional plasmid-- make sure...setting up a small-scale expression experiment to test variables such as temperature, time, and media conditions...temperature or moving it to a bench-top shaker 10-20 minutes before adding the inducer. Growth Media: Changing...

  5. Plasmids 101: The Promoter Region – Let's Go!

    Type
    Blog Post
    ...tightly regulated. Common promoters for eukaryotes and prokaryotes We have included two reference tables...bacteria this is done by a single enzyme; however, eukaryotes have muliple polymerases which are each responsible...expression mRNA Yeast promoter containing Gal4 binding sites, commonly used in Drosophila Specific Requires the...

  6. Four Ways to Package Transgenes That Exceed the Size Limit of Adeno-associated Virus

    Type
    Blog Post
    ...different splice site donor and acceptor sequences were tested and with the optimal sequences, the group achieved...homologous recombination approach delivers large donor templates using two sequential homology directed repairs...2017) CRISPR-Mediated Integration of Large Gene Cassettes Using AAV Donor Vectors. Cell Reports 20:750–756...PA (2002) An mRNA Surveillance Mechanism That Eliminates Transcripts Lacking Termination Codons. Science...

  7. Transferable Skills Guide: Conflict Resolution

    Type
    Blog Post
    ... Fig 1: Quote from Michael Ende, courtesy of morefamousquotes.com A good listener absorbs and remembers...college, when I discovered that many of my old suitemates stopped talking to each other if I was not there... diagram of the Thomas-Kilmann Conflict Model, courtesy of https://managementweekly.org/thomas-kilmann-conflict-resolution-model...

  8. Antibodies 101: Introduction to Immunofluorescence

    Type
    Blog Post
    ...molecules, knowing that where you see fluorescence indicates your target’s location. Some other terms you might... The biggest difference between IF and ICC/IHC relates to the type of secondary antibody you use, so as...corresponding protocols. Other researchers may also have tested an antibody in IF and will share their findings...of interest. Thankfully, there are a number of websites compiling antibody data that can help you compare...

  9. Fluorescent Proteins 101: When GFP lets you down

    Type
    Blog Post
    ..., followed by an intramolecular reaction that generates the chromophore (Tsien, 1998). Importantly, this...absorb light at 488 nm. The ratio between the two states is reflected by the pKa, which is around 6.0 for...relatively small, adding GFP may alter diffusion rates. Smaller genetically encoded probes are based on...Adjobo-Hermans, 2011). Structural information facilitates the choice of the proper site, although it still...

  10. Adeno-associated Viruses (AAVs) for Genome Editing

    Type
    Blog Post
    ...approach is most well suited, since it routinely generates knockin efficiencies of 1-40% with G418-selected... AAV to deliver their repair template to mouse zygotes and thereby correct a mutation that causes Hemophilia...technique to efficiently deliver large repair templates for applications that require large gene fusions...Porteus. "CRISPR-mediated integration of large gene cassettes using AAV donor vectors." Cell reports 20.3 (2017...

  11. Targeting HIV-1 with CRISPR: Shock and Kill or Cut it Out?

    Type
    Blog Post
    ...-target sites with up to 7 mismatches to the gRNA target sequence. Kaminski et al. next tested their procedure...targeting of the enhancer region near the NF-KB binding sites increased activation of HIV promoter-luciferase ...expression of Cas9/gRNAs, which could result in higher rates of off-target cleavage. In both cases, translating...

  12. Plasmids 101: Gibson Assembly and Other Long-Homology Based Cloning Methods

    Type
    Blog Post
    ...not require the presence of specific restriction sites (or lack thereof) nor a serious time commitment....of fragments, but uses site specific restriction sites (Type IIS restriction endonucleases) to cleave DNA...that the vectors and DNA fragments contain these sites at the correct location and NOT in the middle of...flanked by orthogonal versions of the attB and attP sites recognized by the integrase. This method requires...

  13. Generating Mouse Models Using CRISPR/Cas9

    Type
    Blog Post
    ...(Wenning) was fascinated by CRISPR and rushed to test it. I still remember the day, March 16, 2014, that... it has been repurposed for genome editing in eukaryotes, with the most widely used CRISPR genome editing...specifies the target, the Cas9 endonuclease that creates the DNA double-strand break (DSB) at the target...mouse zygote (Qin et al., 2015).  When inside the zygotes, the gRNA will seek out its target among the 3 ...

  14. Building Global Connections with the International Mentorship Program USA-EUROPE

    Type
    Blog Post
    ...country of origin with an international mentor creates a closer and more approachable relationship for...evidence of this, you needn’t look any further than testimonials from the students who participated in the program... of Oviedo, the European University Miguel de Cervantes, the University of Navarra, and the University...macular degeneration. She is also a blogger who writes posts for the blogs of MasScience, ECUSA, IMFAHE...

  15. Antibodies 101: Introduction to Antibodies

    Type
    Blog Post
    ...the surface of extracellular pathogens such as parasites or microbes, or to proteins expressed on the surface...cascades that clear these infections. Anything that generates an antibody response in the immune system is referred...at different times of the immune response and initiates different immune cascades. In the research setting...quantitative measurement of protein expression in cell lysates, whole cells, or tissue samples. Many experiments...

  16. Cloning

    Type
    Guide
    ...enzyme sites. You can easily move (subclone) any piece of DNA that already has restriction sites on either...systems is that like restriction enzyme sites, type IIS sites can be found throughout DNA sequences. Thus...double stranded DNA (dsDNA) at specific sequence sites in the DNA. In restriction cloning, scientists utilize...either side of it into any plasmid that has the same sites in the same orientation within its multiple cloning...short length, it is also easy to add restriction sites to any piece of DNA during PCR amplification, allowing...important to note that restriction enzyme target sites can be repeated throughout a specific DNA sequence...already be surrounded by specific recombination sites (in this regard, not so dissimilar from restriction...

  17. 2021 Deck the Lab Winners!

    Type
    Blog Post
    ... announce the winners of our 2021 Deck the Lab Contest! Though 2021 had more than its fair share of challenges... challenges, the annual Deck the Lab Contest brought us smiles and joy. In fact, our Addgenies were absolutely...festive makeover today✂️❄️🦠thanks to the awesome templates by @CVRHutchinson @CVRinfo. Check them out: https...

  18. Molecular Biology Reference

    Type
    Guide
    ...of DNA, or templates, are spatially separated from each other. These attached DNA templates are then amplified...Alternative start codons include CUG in eukaryotes and GUG in prokaryotes. Common Epitope Tags Tag Amino Acid...DNA. Although they can be found in archaea and eukaryotes, they play the most significant biological role...fragment with a plasmid or vector DNA backbone generates a recombinant DNA molecule, which can be used ...of DNA which contains several restriction enzyme sites, enabling easy insertion of DNA by restriction enzymes...the promoter. As a general rule, the restriction sites in the MCS are unique and not located elsewhere ...relatively quickly, and exhibit exponential growth rates, plasmids can be replicated easily and efficiently...

  19. Antibody Data Repositories and Search Engines

    Type
    Blog Post
    ... start looking to find one. Luckily, there are websites that can help: Antibody data repositories, which... Website Targets Application Purpose Notes Antibodypedia Any Any Data repository ...Molecules group Addgene Data Hub Any (see notes) Any Data repository Only accepts data for...

  20. Countdown to Halloween @Addgene

    Type
    Blog Post
    ...through 2009, though I suspect that our costume contests were indeed occurring earlier but that the photo...protect the innocent revelers. Our Halloween costume contests have become an annual tradition in which we choose...finding out who wins this year’s Addgene costume contest, like us on Facebook and checkout our FB page on...
Showing: 461 - 480 of 963 results