We narrowed to 978 results for: LARS
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TypeBlog Post...cell type? You can use DNA sequencing or other molecular cloning techniques to determine CRISPR/sgRNA efficiency...site in the genome you are trying to target play a large role in Cas9 activity and efficiency (Sander and... structure and genomic recognition sites play a large role in CRISPR-Cas9 efficiency. They continued to...
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Synthesized by Ginkgo Bioworks, Shared by Addgene: SARS-CoV-2 Plasmids for Many Expression Systems
TypeBlog Post...genes, tags, truncations, and codon optimizations similar to those found in the bacterial expression plasmids...Fukushi S, Watanabe R, Taguchi F (2008) Pseudotyped Vesicular Stomatitis Virus for Analysis of Virus Entry Mediated...SARS Coronavirus Spike Proteins. In: Methods in Molecular Biology. Humana Press, pp 331–338 https://doi.... -
Antibodies 101: Flow Cytometry Controls
TypeBlog Post...unsure what your population will look like for a particular marker, you can use the reference plots provided...isotype controls An isotype sample is prepared very similarly to a FMO sample: you make a sample from a combined...and then add in your isotype control for that particular marker. This enables you to identify any non-... -
The Many Reasons for Irreproducible Research- and a Vaccine to Eradicate It
TypeBlog Post...and not just the subset of those that return a particular result. Anything else that occurs during the ...are known. The author submits the research plan (similar to a preregistration, but usually with a fully ...Promotion Guidelines; preregistration to increase clarity in study design and analysis; removing publication... -
A Novice's Perspective on Neuroscience - SfN2017
TypeBlog Post... almost all of the attendees were going to be molecular biologists. This was not at all the case. The ...neurobiology (and really in all of biology) require large data sets. Whether these data sets include videos... make the virus for you. Neuroscientists are particularly interested in using AAVs because they provide... -
Top Requested Lentivirus and AAV of 2016
TypeBlog Post...used to investigate the effects of activating particular signaling pathways in specific cell types (typically...typically neurons). While all of these viruses are popular, pAAV-hSyn-DIO-hM4D(Gi)-mCherry contains a double...animals, where cre expression is restricted to a particular cell type. The human synapsin promoter further... -
Back to Bacteria: CRISPR gRNA Multiplexing Using tRNAs
TypeBlog Post..., gRNA and pol III terminator. These relatively large cassettes (considering the gRNA itself is ~100 bases...advantageous to instead use a preexisting, conserved cellular machinery to process gRNAs. Hijacking tRNA processing...frequency of both indels at each locus, as well as large deletions. They detected fragment deletion with ... -
How to Make Friends and Meet People at a Scientific Conference
TypeBlog Post...national recognition and the more people you know, the larger your profile in the community Job hunting outside...thing to consider is the size of the conference. Large conferences have many opportunities but can be overwhelming... overwhelming as far as meeting new people. At large conferences, look for smaller events or meet-ups ... -
Summer SciComm Series: Cognitive Load
TypeBlog Post...bandwidth. Jargon Learning, using, and retaining new vocabulary takes time and effort (otherwise it would be ...achieve!) Each field has its own framework: a particular way of organizing and connecting information ...experiences. If you’re hearing multiple people with similar misconceptions, it’s time to rework your strategy... -
Transferable Skills Guide: Problem Solving
TypeBlog Post...Support Team here at Addgene, so troubleshooting molecular biology experiments was in the job description...fuzzy or ill-defined. In this article, I try to clarify what people mean when they ask about “problem solving...result of resolving the issue. Depending on the particular problem, you might even be expected to write ... -
Tagging Optogenetics and Chemogenetics Receptors: Fluorescent Proteins and Other Options
TypeBlog Post...microscope showed us that, in monkey neurons, the large majority of the gold particles were in the cytoplasm...after it has been administered. Although we found a larger proportion on the membrane in mice neurons, most...the same cell, but not necessarily at the same subcellular compartment as the opsin or chemogenetic receptor... -
Stabilized Bacterial Promoters: Constant Gene Expression at any Copy Number
TypeBlog Post...or other synthetic biology circuits. What is a molecular biologist to do? Let’s take a look at how plasmids...expression from the constitutive promoter did not. Similar levels of expression between a plasmid and the ...stabilized promoter, deoxychromoviridans titers were similar regardless of where the genes were expressed. What... -
Celebrating 15 Years of Scientific Sharing
TypeBlog Post...worry about getting support from depositors. We regularly hear how much they see the value in depositing...) and even on 60 Minutes. We are blessed with a large community of scientists who support us by depositing... I checked we were ranked as the 21,806th most popular website in the world. I know, right? Upgrades to... -
The time and cost required to make a plasmid
TypeBlog Post... repositories like Addgene provides large-scale savings. How large? Using Doench’s calculations and the...purchases can raise the cost anywhere from a few dollars to over a hundred. Of course, these were all 2018... -
RNA Extraction Without A Kit
TypeBlog Post...Check out our molecular biology protocols Find protocol videos on our website Read our Molecular Biology Guide...many samples you may have, access to one or more large, expensive ultracentrifuge. Figure 1: An ... -
Hot Plasmids - March 2020 - base editors, GEVI, MoClo, and optogenetics
TypeBlog Post...packaging capacity of 4-5 kb limits the delivery of larger transgenes, such as base editors which have a gene...efficiency, the split editors transduced at levels similar to that of a single AAV expressing GFP. The lab...cyanobacteria Article contributed by Andrew Hempstead The Modular Cloning (MoClo) system allows synthetic biologists... -
Pairing CombiGEM and CRISPR for Combinatorial Genetic Screening
TypeBlog Post...CRISPR guide RNAs (gRNAs) for genome editing in large-scale has led to the rapid generation of genome-...combinatorially modify the genome, screen for a particular phenotype, and quickly profile the resultant ...Sciences of the University of Hong Kong. He is particularly interested in developing and applying new technologies... -
Nanoblades: Tiny CRISPR Ninjas for Genome Editing Difficult Cells
TypeBlog Post...sequence ranging from a single nucleotide change to large insertions like the addition of a fluorophore or...components. However, this sometimes requires intracellular injections into the cytoplasm or pronucleus ... albino mice ranging from 11% to 100%, a level similar to other approaches. Nanoblades can also be used... -
CRISPR 101: Anti-CRISPR Proteins Switch Off CRISPR-Cas Systems
TypeBlog Post...., 2020). Some Acr proteins are specific to a particular Cas protein while others can inhibit CRISPR enzymes...can increase the chance of off-target editing particularly after the majority of on-target editing has ...Cas9 Toxicity in Human Hematopoietic Stem Cells. Molecular Therapy - Methods & Clinical Development 9:390... -
Cpf1 Update: Comparison to Cas9 and NgAgo
TypeBlog Post.... Sticky-end mediated gene transfer would be particularly helpful for targeting non-dividing cells, which... are 3.7 kb and 3.9 kb in length, respectively, larger than SaCas9 but smaller than SpCas9. These nucleases...et al. suggest that AsCpf1’s off-target rate is similar to that of high fidelity Cas9s eSpCas9 and SpCas9...