...as CRISPR-based gene repression tools to find and test genes that may play a role in the development of...polymerases produced using plasmids from Addgene to test for the presence of pathogens in environmental samples...

...Golden Gate cloning method to ligate expression cassettes of the gRNAs into a single vector in tandem. They...nickase vectors work successfully to target multiple sites in cultured human cells. While it has not yet been...

...be worn at all times. Use plastic pipettes in place of glass pipettes or needles. Liquid waste should be...shRNA oligos are cloned into the AgeI and EcoRI sites in place of the stuffer. The AgeI site is destroyed... 10x NEB buffer 2 35 μL ddH 2 O Incubate for 4 minutes at 95°C in a PCR machine or in a beaker of boiling...PCR machine, incubate the sample at 70°C for 10 minutes then slowly cool to room temperature over the period...following manufacturer’s protocol. Plate on LB agar plates containing 100 μg/mL ampicillin or carbenicillin...swirling or gently flicking the tube. Incubate for 5 minutes at room temperature. e. Add 80 μL of FuGENE® master...gently flicking the tube. f. Incubate for 20-30 minutes at room temperature. g. Retrieve HEK-293T cells...

...demonstrate efficient editing in adult mice and in zygotes. Read the paper in Molecular Cell Find the Nme2Cas9... was developed by Rebecca Shapiro’s lab. After testing dCas9 fusions with transcriptional repressors, ...

...are used to insert recombinant DNA from prokaryotes or eukaryotes into a synthetic vector that can have ...

...2015 was... lentiCRISPR V2   LentiCRISPR v2 originates from Feng Zhang’s lab at the Broad Institute ...secretion, and identifying lysine methyltransferase substrates. We’re incredibly happy that lentiCRISPR v2 is...

... This blog was originally published on BitesizeBio here. As part of my job ensuring plasmid quality at...Verify Plasmids Acknowledgements: Thank you BitesizeBio for originally publishing this and allowing us...

...nature of the Center for Regenerative Medicine accelerates research. Chapter 5: Research currently under... defines Open Science and how he practices and promotes open science in the lab - sharing reagents, cell...

...be repurposed for double strand breaks at target sites. Read the paper in Nature Communications Find...is a single step gRNA multiplexing system that generates vectors containing high numbers of gRNAs in one...

...And could it be a new tool for genome editing? To test the potential of Cas14, the Doudna lab cloned the...of dsDNA and RNA in trans. Fluorescent signal indicates the presence of this viral sequence in the assay...

...compensation committee. The compensation committee creates a compensation philosophy that helps guide employee...spend time caring for my mother. For fun, I enjoy pilates, jewelry making, reading, sailing, and gardening...

...transfers. The UBMTA was drafted by the National Institutes of Health (NIH) and adopted by the Association... Processing: When you place an order, Addgene generates the MTA and emails it straight to you, or we can...

...being plated on agar with ampicillin selection.  Notes and tips  We’ve found that our competent cells ...contamination.  Also conduct a positive control (test a plasmid that transforms well in other competent...

...DNA for 2 minutes at 72°C. Repeat steps 8-10 for 25-30 cycles. Final Extension for 5 minutes at 72°C. ...oligo primers. This molecular biology technique creates several micrograms of target DNA from just a few.... Basic PCR Program Initial Denaturation for 2 minutes at 94°C: This initiation step heats the double ...Repeat steps 2-4 25-30 times. Final Extension for 5 minutes at 72°C: A final extension to fill-in any protruding...melting temperature (Tm). Set extension step at 1-2 minutes per kilobase of product depending on whether you...time and temperatures. Initial Denaturation for 2 minutes at 94°C. Denature for 30 seconds at 94°C. Anneal...denaturation of the DNA, particularly for GC-rich templates. What does each ingredient specifically do? Template...

...Michael Koeris to learn how his company uses phage to test for Listeria in food products). I picked this post...Plasmids at a Low Price: Kits from Addgene Savvy Advocates Needed to Navigate a Scientific Enterprise in ...

...might have a few positive control genes on which to test the validity of your biological model and the technical...starting point for developing a model and an assay to test that model. Both libraries are available from Addgene...

...which labs should receive the clinical sample to test their phages. Because the sample is limited, only...highest potential to possess the correct phages will test. These includes labs with a large number of different...

...save you the time that would be spent making and testing entirely new gRNA designs. You can now easily find...variations (e.g. SNPs) exist. Additionally, consider testing multiple gRNAs targeted to the same locus to ensure...

...isolation of successful transformants. Addgene distributes several ready-to-use, modular plasmids, combining...combining fluorescent tags, epitope tags, protease sites, and selection markers. These are especially useful...

...research by anyone. As infection rates were increasing in the United States, and many universities and Technology...