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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 978 results for: LARS

Showing: 681 - 700 of 978 results

  1. The PAM Requirement and Expanding CRISPR Beyond SpCas9

    Type
    Blog Post
    ...are not always positioned correctly to target a particular gene. Furthermore, a target sequence may have... desired target locus. The PAM sequence is of particular concern when trying to edit a gene using homology...cleavage events for the VQR and VRER variants is similar to wild-type SpCas9, indicating that the variants...

  2. Plasmids 101: Broad Host Range Plasmids

    Type
    Blog Post
    ...known to have replicons with broad host ranges. Popular replicons from these groups include: RK2 (IncP)...vectors for gene expression in bacteria. Methods in Molecular Biology (Clifton, N.J.), 765, 327–343. https:/...endonucleases on the broad host-range plasmid RK2. Molecular & General Genetics: MGG, 152(3), 129–135. https...

  3. Plasmids 101: Control Plasmids

    Type
    Blog Post
    ...level of Gene X in cells treated with Plasmid A is similar to the expression level of Gene X in the untreated... controls are negative controls that produce a similar product, but do not target an endogenous gene in... variable and helps ensure the phenotype isn’t singular or associated with only one aliquot/preparation...

  4. Protocol - How to Run an Agarose Gel

    Type
    Protocol
    ... the gel in the first place. Carefully load a molecular weight ladder into the first lane of the gel. ...you are conducting certain procedures, such as molecular cloning, you will need to purify the DNA away ...you get better separation of bands? If you have similarly sized bands that are running too close together..., and a lower percentage gel will help separate larger bands. 10% Rule For each sample you want to load...

  5. What is Polymerase Chain Reaction (PCR)

    Type
    Protocol
    ...cytosine (C), guanine (G), thymine (T)] in an equal molar ratio. It also requires two unique single stranded...copy the DNA in between the oligo primers. This molecular biology technique creates several micrograms of...Your 5’ and 3’ primers should be designed to have similar melting temperatures (Tm). Set the annealing temperature...frozen. DMSO helps with denaturation of the DNA, particularly for GC-rich templates. What does each ingredient...

  6. Tips for Getting a Faculty Position

    Type
    Blog Post
    ... matter how many times I say this, candidates regularly try to convince me that their search committee...whiskies, but I remember the days of modest postdoc salaries all too well. If there's a group to make money...search!   Erik Lee Snapp received his PhD in Molecular Microbiology and Immunology from Oregon Health...

  7. Deep Mutational Scanning with One Pot Saturation Mutagenesis

    Type
    Blog Post
    ...functional screen of your liking. This method is similar to site-directed mutagenesis (SDM) PCR, but with...Random Mutagenesis by Error-Prone PCR”. Methods in Molecular Biology In Vitro Mutagenesis Protocols, pg. 103...Measuring the Activity of Protein Variants on a Large Scale Using Deep Mutational Scanning.” Nature Protocols...

  8. 10 Steps to a Perfect Science Talk

    Type
    Blog Post
    ..., but I always seek out information about the particular audience and venue before tailoring each presentation...peer mentors to check your slides for typos and clarity in a big room.  Finally, don’t pass up opportunities...informal poll by asking for a show of hands on a particular question. I like to ask about how many people...

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

    Type
    Blog Post
    ...efficiency differed from 48-100% between strains. In a similar experiment, CRISPR reduced the HIV-1 copy number...evidence that an HIV-1 cure may be within our reach. Similar studies have shown that CRISPR can be used to combat...Additional Resources on the Addgene Blog Treating Muscular Dystrophy with CRISPR Gene Editing Learn to Use...

  10. Coomassie Purity Stain of Recombinant Antibodies

    Type
    Protocol
    ...and reagents from other vendors should produce similar outcomes when using these protocols. However, please...blade Plastic tray Gel imaging system ImageJ or similar software Reagents Purified recombinant antibody...should be very little background staining. Pro-Tip A large shift in the electrophoretic mobility of the bands...Analysis Use ImageJ (Link opens in a new window) or a similar photo software to determine the relative intensity...

  11. How to Design Your gRNA for CRISPR Genome Editing

    Type
    Blog Post
    ... to choose from that sequence preferences must largely be ignored. Created with BioRender.com. ...nuclease-mediated off-target effects.” Further, large-scale datasets of hundreds of genetic screens using...Tsherniak A, Iorio F (2019) Agreement between two large pan-cancer CRISPR-Cas9 gene dependency data sets...

  12. Antibodies 101: Multiplex Immunofluorescence

    Type
    Blog Post
    ...appropriate secondary antibodies. The general process is similar to standard immunofluorescence, but with some extra...selecting an antibody, some of the requirements are similar to standard IF. Here, we’ll walk you through the...if the localization and intensity patterns are similar between the two experiments. If not, it might be...

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

    Type
    Blog Post
    ...Mentorship Program work? IMP Pilot Program 2015-2015 Scholars need strong proactive mentors who have relevant...President from the non-profit IMFAHE, Inc. She is particularly interested in breast cancer and metastasis as... Medical School, where she studies age-related macular degeneration. She is also a blogger who writes ...

  14. AAV ddPCR Titration

    Type
    Protocol
    ... Service Viral Vector Guides Virus Blog Posts Molecular Biology Reference Introduction This protocol describes... lower or higher dilutions depending on their particular sample. For additional tips on AAV titering using... bucket 96-well freezer blocks (x 3) Reagents Molecular Biology Grade Water, Hyclone, SH30538.02 GeneAmp...10X Buffer 25 µL of 10% Poloxamer 188 4475 µL Molecular Biology Grade Water Procedure Preparation Before...

  15. 9 tips for a successful postdoctoral experience

    Type
    Blog Post
    ...reason most people enter postdoctoral training. Similarly, there are several factors to consider when selecting...to strengthen your new model?  PFs need to be regularly thinking about manuscripts. The other items will...Erik Snapp! Erik Lee Snapp received his PhD in Molecular Microbiology and Immunology from Oregon Health...

  16. Protocol - How to Purify DNA from an Agarose Gel

    Type
    Protocol
    ...samples. This is a commonly used technique for molecular cloning, such as PCR - or restriction enzyme -...you get better separation of bands? If you have similarly sized bands that are running too close together..., and a lower percentage gel will help separate larger bands. 10% Rule For each sample you want to load...

  17. Protocol - How to Perform a Diagnostic Digest

    Type
    Protocol
    ...result in similar sized bands, thus making this simple digest less informative. This is particularly true ...occurring bacterial endonucleases that recognize a large range of DNA sequences. Given the variety of these...

  18. Protocol - How to Inoculate a Bacterial Culture

    Type
    Protocol
    ...usually want to start 2 mL in a falcon tube, but for larger preps you might want to use as much as a liter ...individual plasmid within a single bacterial cell. Large plasmids usually have a low copy number (approximately... other hand, smaller plasmids can be present in large numbers, 50 or more per cell and have a high copy...

  19. Protocol - Bacterial Transformation

    Type
    Protocol
    ...less efficient at taking up larger plasmids. If you need to transform large plasmids, it is a good idea...very few if any transformants when transforming large plasmids (>10 kb) or BACs, what can I do? Chemically...
Showing: 681 - 700 of 978 results