...during Zoom lab meetings to chip in ideas and take minutes that can be accessed by anyone at any time. This... be written by several authors from different institutes, disciplines, and cultural backgrounds.” This...simply moves away from the desk to switch off a few minutes. The host could also set aside Breakout Rooms to... allowed with a different host each time that moderates conversations. This can help team building and...

...In addition to single plasmids, Addgene also distributes pooled plasmid libraries containing hundreds,...can be counteracted by growing the cells on solid plates, where each transformant forms its own colony, ...solid media. The colonies are then scraped off the plates and pelleted for DNA extraction. Prepare library...using both pre- and post-amplification DNA and generates PCR sequencing products from both. These products...

...dissociation rates and discovered that the catalytically active form of the nuclease dissociates very slowly...our surprise, we measured identical dissociation rates for catalytically inactive dCas9 molecules. We followed...precisely spaced region allowed sequence replacement rates of approximately 1%. This is by no means a large...

...is a challenge. To help keep you abreast of the latest fluorescent proteins, I maintain an interactive...match to the filter set but the excitation filter excites more of the peak of the TagRFP excitation and the...to be extremely bright. In S. cerevisiae, we've tested a number of green and red fluorescent proteins ...optimal for all organisms and that if you want the brightest signal, you may need to try several proteins in...

...control AAV in smaller aliquots of 20ul for serotype testing to help you pick the best AAV for your experiment...GABA-ergic interneurons (83895). See our Serotype Testing AAV inventory for the entire collection! Plasmid...activated, this DREADD acts through the Gq pathway and promotes neuronal firing. Plasmid Serotype Name Depositor...functional Cre enzyme that induces recombination of lox sites. This system takes advantage of the extensive supply...

...none of these constructs have been functionally tested by Ginkgo Bioworks or Addgene. If you used plasmids...combinations of WT, codon optimized, untagged, cleavage sites (thrombin and TEV), and C-terminal tags (StrepII...first pass QC. The lab team manually transforms, innoculates, and performs DNA minipreps for small batches...mutation.  Once the samples arrive, each group of plates will have select samples prepared and screened ...

... PAM specificity, allowing for the targeting of sites that were previously unavailable. Figure ... with a broader PAM specificity can target more sites. Created with BioRender.com. Specificity Specificity...variants that were able to target different PAM sites (NGG, NGAN, NGNG, NGAG, and NGCG). Since then, the...eSpCas9 and SpCas9-HF across a number of different sites. Using a combined random and directed mutagenesis...

...of Producing Monoclonal Antibodies, 1999). This creates an immortal cell population that will continue ...of molecules including proteins, lipids, and carbohydrates, in this blog post we’ll focus on antibodies...lots over time. Specifically, the antigen-binding sites of the antibody - the paratopes - may change and... Addgene’s antibody plasmid collection Read the latest on Addgene's new antibody project ...

...cleavage, and suppresses neuronal transmission within minutes. Constructs using the photodimerizer system CRY2...Escherichia coli and Salmonella Typhimurium. This work demonstrates engineered gas vesicles as the first reporter...is regulated in the 3’ UTR by four miRNA target sites and thereby serves as a sensor of miRNA presence...population, or identify differentiation and disease states. This collection of reporter plasmids is amenable...

... 49,000 times! Their long-lasting popularity illustrates the utility these tools have in many different...pyogenes with an invariant sgRNA scaffold and cloning sites for inserting the guide sequence (pSpCas9(BB)). ...notice the “V2.0” at the end of this plasmid which indicates that there was a prior version of this plasmid...retroviral). Fun fact: published in 2003, this plasmid predates Addgene’s founding. Find pCMV-VSV-G. pRSV-Rev...

...These small, but mighty type VI-D enzymes are the latest tools in the transcriptome engineering toolbox....sequence! In bacteria, Cas13d-mediated cleavage promotes collateral cleavage of other RNAs. As with other...efficiency varies widely across guide RNAs, each guide tested with CasRx yielded >80% knockdown. It seems that...maintains its RNA-binding properties, Konermann et al. tested its ability to manipulate RNA species through exon...

... want to buy, it is worthwhile to test-drive a couple of candidates. Before you buy a new microscope, ...summarizes properties of fluorescent proteins on websites. To make the best choice, the fluorescent protein...than theoretical brightness for selecting the brightest fluorescent protein. Theoretical brightness Two...

...or PolyPhen to approach this challenge, using attributes of cancer variants such as evolutionary conservation...have been two huge challenges preventing us from testing this hypothesis: There was no large scale, broadly...whether gene-agnostic bioassays could accurately test for molecular signatures of variant function and...the second major challenge, we developed assays to test large numbers of cancer variants across a large ...

...fantastic K12 education initiatives like BiteScis (https://bitescis.org/) and amazing graduate student writing...writing collaboratives like Oceanbites (https://oceanbites.org/) have already grown out of our workshops...

...finishes repair of most types of breaks in tens of minutes – an order of magnitude faster than HDR. NHEJ is...paired end complex The paired end complex then ligates compatible DNA ends together, thus repairing the...Aprataxin, Tdp2) that function to make ends better substrates for ligation. Although we do not describe these...vastly reduces breaks and mutations at off-target sites. Christina Mork contributed to the update of this...

...relevant to the disease in question – cardiomyocytes, adipocytes or neurons, for instance – to see how...Cell Stem Cell paper, for example, we generated hepatocytes to examine the effect of a gene on cholesterol...Musunuru: The CRISPR/Cas9 system is easier to use as testified by the fact that the TALEN kit includes 834 plasmids...

.... ANOVA, t-test, multiple regression analysis, and non-parametric analogous Wilcoxon tests have all been...variation into your data. Try different types of pipettes and pipetting methods to find what’s most consistent...plate. Compare the CT values within each set of triplicates to understand variation due to technical error...reproducibility’s sake!   Fig. 2: Setting up triplicates in multiple configurations can help you identify...

...splitting removes any exogenous sequences and regenerates a native peptide bond at the split site. ...to that of a single AAV expressing GFP. The lab tested the base editors in vivo and achieved therapeutically...used in human gene therapy trials. They further tested the base editors by correcting a mutation that ...synthetic biologists to use Type IIS restriction sites to easily assemble multiple DNA fragments. This ...

...these plates now” to a fellow team member. But, at Addgene, we use seven different types of “plates” (Fig... meetings. A lot of our LIMS planning involved “plates,” and our scientists and lab technicians always...information Prescribing an official name to all of our plates, tubes, and racks meant we had fewer errors in ...

...the accumulation of toxic intermediates or the shuttling of the intermediates into other pathways. In cases...of plasmids, the stabilized promoters were also tested in different plasmid backbones that varied in size...gene copy number is affected differently by growth rates when a gene is expressed from a plasmid as opposed...