We narrowed to 526 results for: erf
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TypeBlog PostPublishedOct. 13, 2022, 1:45 p.m....Addgene’s repository) and make it easy to find your perfect match. Choosing a Cas9 for your knockout experiment...deaminase to where the edit is desired, and it performs the programmed edit within the editing window ...with this tool is that the desired edit will be performed on all relevant bases within the editing window...
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Avoiding the Dark Side of Fluorescent Protein Fusions with mOX FPs
TypeBlog PostPublishedOct. 27, 2015, 3 p.m....cannot be ignored. They have the potential to interfere with the function of resident ER chaperones and...and retained in the ER. FPs have been undeniably powerful and successful tools that have enabled numerous..., Lindsey M. Costantini, and Erik L. Snapp. "Superfolder GFP is fluorescent in oxidizing environments ... -
Plasmids 101: Control Plasmids
TypeBlog PostPublishedApril 9, 2015, 3:29 p.m.... to the consistency of the reagents and hands performing the experiment, not the reproducibility or consistency...independent aliquots of cells and plasmids, be performed on different days, etc, but outside constraints...incorporating additional positive and negative controls, performing multiple replicates, and ultimately getting interpretable... -
Antibodies 101: Beyond Surface Labeling
TypeBlog PostPublishedJune 27, 2024, 1:15 p.m....mummification). Permeabilization, as the name suggests, perforates the cell membrane to allow for antibodies to ...day. Note that intracellular labeling must be performed only after viability staining and surface labeling...surface labeling. Intracellular labeling can be performed on cytosolic and/or nuclear markers. Lastly, stains... -
CRISPR 101: Validating Your Genome Edit
TypeBlog PostPublishedNov. 3, 2022, 12:15 p.m....analysis is performed yielding a graph of edits in the input population or clone. How to perform a TIDE...homology-directed repair events, or maybe you’re performing a single base pair edit with a prime editor. ... -
SARS-CoV-2/COVID-19 Detection Methods Based on CRISPR/Cas
TypeBlog PostPublishedMay 5, 2020, 1:15 p.m.... virus directly using saliva without having to perform RNA extraction but that may work only if there ... Motif) for target cleavage. This test can be performed in two steps for challenging samples as in Two-Step-SHERLOCK...fluorescently labelled substrate, gets activated and performs non-specific collateral cleavage. This cleavage... -
Fluorescent Proteins 101: When GFP lets you down
TypeBlog PostPublishedAug. 23, 2018, 12:05 p.m.... GFP is such a well-validated, all-round good performing probe, it is the first choice when selecting ...reduces its size. The substantial size of GFP may interfere with the activity or the localization of the fusion...C-terminus of another protein. This may, however, interfere with the function of the protein. This is clearly... -
Antibodies 101: Multiplex Immunofluorescence
TypeBlog PostPublishedMay 23, 2023, 1:15 p.m....repeating for each target. Alternatively, you can perform simultaneous detection with a mixture of primary...steps, in order to ensure your antibodies are performing their best and that the sample is not being degraded... P., & Yong, W. H. (2019). An Introduction to Performing Immunofluorescence Staining. In W. H. Yong (Ed... -
Plasmids 101: Dimers and Multimers
TypeBlog PostPublishedJan. 10, 2023, 2:15 p.m....When performing restriction digests on plasmids at Addgene, we sometimes observe something odd in our...Running undigested DNA on a gel as a control when performing a diagnostic digest is a simple but effective... sequencing Addgene uses Illumina MiSeq NGS to perform whole plasmid sequencing of our plasmid samples... -
Easi-CRISPR: Generating Knock-In and Conditional Mouse Models
TypeBlog PostPublishedApril 5, 2018, 12:42 p.m....T7 promoter upstream of the left homology arm, perform in vitro transcription, and use the IVT product... and infundibulum. In vivo electroporation is performed immediately after injection. Image from Ohtsuka...contrast to standard microinjection, i-GONAD is performed directly on pregnant female mice using a small... -
PiggyBac-ing Through the Genome Editing Field
TypeBlog PostPublishedMay 31, 2016, 3:30 p.m....can be combined with CRISPR/Cas9 to efficiently perform genetic manipulations in animal models or cells...activity. In summary, the piggyBac™ transposon is a powerful technology that may be used for a variety of research...piggyBac™ for gene editing allows you to efficiently perform seamless genetic manipulations, while avoiding ... -
Plasmids 101: Restriction Cloning
TypeBlog PostPublishedFeb. 18, 2016, 3:42 p.m....Read on for an in-depth breakdown of how to do perform restriction digests. Overview of the restriction... common chemically-competent cells. You should perform, at minimum, two transformations after a ligation... 101 Posts Resources on Addgene.org Find the Perfect Empty Vector for Your Cloning Experiments Browse... -
Tips for a 1st Time CRISPR User (by a 1st Time CRISPR User)
TypeBlog PostPublishedMarch 7, 2017, 3:30 p.m....what I think are the main tips for each step of performing and analyzing a gene edit using Addgene’s lentiviral...lentiCas9-blast. Anti-Cas9 Western blotting was performed on whole-cell extracts from stable pools of A549...blasticidin selection. Anti-Cas9 Western blotting was performed on whole-cell extracts from the initial stable... -
An “elegans” Approach to Better CRISPR/Cas9 Editing Efficiency
TypeBlog PostPublishedDec. 17, 2020, 1:30 p.m....phenotypes. These approaches may become even more powerful following a report that one can use linear repair...non-linked loci; pha-1(e2123) mutant worms are perfectly viable at 15 ºC, yet exhibit complete embryonic...recombination repair pathway. Reagents required to perform pha-1 co-conversion are available through Addgene... -
Finding Your Science Policy Path
TypeBlog PostPublishedSept. 22, 2020, 1:15 p.m....and grow along the way. That’s what makes it so wonderful. The most challenging part of jumping into this...me, the Veterans Health Administration was the perfect position because my science policy ‘why’ was to...this question may change over time and that’s perfectly normal! Action Items: Start a career journal... -
CRISPR Methods for Bacteria: Genome Engineering, CRISPRa, CRISPRi, Base Editing, and More
TypeBlog PostPublishedSept. 28, 2020, noon...homologous recombination has not occurred. This powerful negative selection system ensures high editing...Transcriptional repression (CRISPRi) in bacteria As RNA interference does not function in bacteria, most efforts ...inMycobacterium tuberculosisusing an efficient CRISPR interference system. Nucleic Acids Res 44:e143–e143 . https... -
Addgene’s GIPHY Page is Here to Help You Express Yourself (and Your Gene of Interest)
TypeBlog PostPublishedAug. 17, 2021, 1:15 p.m....“addgene” into the GIF search bar to find that perfect response GIF. We have GIFs for every...via GIPHY If you’re also looking for that perfect GIF for your TikTok or Instagram story, we also... -
Find and Share AAV Data with Addgene's New AAV Data Hub
TypeBlog PostPublishedOct. 8, 2019, 12:33 p.m....long the scientist waited after injection before performing the experiment. Every entry has an accompanying... so that you can see for yourself how the AAV performed in that experimental setup. For example, this ... -
Grad School Advice Part 2: Building Community
TypeBlog PostPublishedOct. 6, 2016, 2:30 p.m....up a performance Ben and Niroshi organized for Tyler's (the host's) thesis defense. This wonderful piece... -
Microbes: Look, Listen, and Tell
TypeBlog PostPublishedAug. 28, 2018, 12:04 p.m....on the floor outside the lecture hall. It was wonderful to see the ad-hoc ‘sciencing’ when Dr. Prakash... By connecting art and science, a fascinating interface is created that produces loads of inspiration ...