...methods avoid many of the drawbacks of kits and typically follow the same principles: cells are lysed, RNAs...method described in Kim and Morrison is nearly identical to that of commercially available kits but instead...B., & Gillespie, D.A. (1979). Preparative and analytical purification of DNA from agarose. Proceedings...

...culprit for these unusual cases was either 1) a physiological effect of SARS-CoV-2 toward destabilizing one...Adenovirus (and/or to its partner, AAV) or 2) a sociological effect of the COVID-19 lockdowns causing a lack...Adenoviruses: General Features. In Reference Module in Biomedical Sciences. Elsevier. https://doi.org/10.1016/B978...

... familial aspect of Addgene. As our size has drastically increased, it would have been a tight squeeze...event! Addgene company culture At Addgene we organically developed the philosophy of  “work hard, play...department) right in your breakroom. 5. Throw some physical activity into the mix It’s easy to sit around ...

...community to help Involve family. If you don’t have local family to help, create “fake” family by forming ...help you do your job better. #8 Thou shalt enthusiastically foster independence Your partner can make ... fold their laundry, take public transportation, call a cab and remember their own school assignments,...

...writing are key skills. Other fields, often highly technical or skills-based ones, may not use them very often...questions about your application. These could be logistical, like briefly explaining a gap in your job history...letter needs to be coherent and free of typos and grammatical errors. It does not need to be a literary masterpiece...

...generate a dot plot (Figure 1). Clustered events are called a population.                             ...of markers you use to define your population is called your “gating strategy.” This series of markers ...around clusters to define populations. This shape is called a ‘gate’; it tells the machine that everything ...

...uses a “nicking” Cas9, in which one of Cas9’s two critical enzymatic residues has been converted to an alanine...deaminases edit A-to-G.) In short, the impaired Cas9 localizes the deaminase to where the edit is desired, and...to dCas9.  This technique has been exploited to localize factors such as transcriptional activators and...

...are subject to excision and ssDNA cleavage by canonical base excision repair enzymes. At the same time...targeted by a cytidine deaminase base DNA editor. Specifically the BFP mutant (BFPH66) contains a histidine...markers, and other enzymes such as Cre-recombinase drastically expanding the base-editing reporter toolbox. ...

...by Joe James from Binning Singletons. The sheer scale of a large conference can be intimidating. And it...when we bring all the bins together at a mixer. We call this Quorum Sensing, after the phenomenon in bacteria... career scientists, students, and those from historically underrepresented groups. Through such mentoring...

...SpCas9 and SaCas9, Zhang’s group used an approach called BLESS (direct in situ breaks labeling, enrichment...post-injection. Livers from these mice were histologically normal and liver injury markers were not increased...enough to deliver with AAV. This system fuses catalytically dead dCas13b to the catalytic domain of RNA ...

... D1135E variant is far more selective for the canonical S. pyogenes PAM sequence NGG compared to wild-...novel CRISPR proteins has and will continue to dramatically increase the number of CRISPR applications. ...Wolfe SA, Sontheimer EJ (2018) NmeCas9 is an intrinsically high-fidelity genome-editing platform. Genome...

...below. The GUS gene (β-glucuronidase) with histochemical GUS-staining is a very useful visual reporter...protein of interest to elucidate the subcellular localization of your protein of interest. Find all plasmids...see Figure 3). This method is probably the most scalable method. Transfer your plant into the dark 16 ...

... relative to empty particles (black arrowhead). Scale bar = 100 nm.   AAV resources Browse AAV preps... a concentration step prior to being frozen. Specifically, lentiviral particles are collected from the... strain of E. coli. In addition, plasmids are typically prepared using endotoxin-free plasmid purification...

... to synthesize a large volume of information: basically, a scientist’s tool kit. I wrote a review article... was cheating. Eventually I learned that it’s practical because there simply isn’t enough time to read...used both. Curate and present some useful data Typically lists and pictures are the most useful parts of...

...blocking off time for different tasks. I use google calendar for this and try to give myself flexibility by...activities that are needed to accomplish these goals. Practically speaking, this means I write everything in a ...down? It’s more viscerally appealing for me to physically cross things out (it just feels good) and I remember...

...clones from that population. You can additionally calculate the minimum number of clones you would have to...consider using a version of Cas9 that has been specifically engineered for reduced off target editing, such...Resources at Addgene: Read our CRISPRGuide Get practical help from our CRISPR Protocols Find gRNA Design...

...even when larger vector genomes are packaged, the physical size of the genomes is still ~4.7kb. But despite...overlapping strategy has a region of homology, typically ~400-1400 bp long between part A and B of the ...overlapping split vector had an improvement in muscle physiological performance.  2. Trans-splicing Overview: For...

...here). Cas9 is commonly tagged with a nuclear localization sequence to enhance targeting to the nucleus...interest (GOI) into plant cells. A) The system typically consists of a Ti plasmid, into which the GOI is...(2020) Targeted delivery of nanomaterials with chemical cargoes in plants enabled by a biorecognition ...

...used to mutate the PAM sequence (an NGG sequence critical for Cas9 cleavage), thereby rendering the resulting...unique sites and allows for the introduction of specifically designed mutations (not limited to frame shift...temperature in the polymerase chain reaction." Analytical Biochemistry 209.2 (1993): 284-290. PubMed PMID...

...vitro, and what obstacles must be overcome for clinical success. ART and HIV-1 reservoirs HIV-1 infects...these methods represent exciting advances in preclinical HIV-1 research that should be pursued in animal... field, the lessons learned in pursuing one technological application can benefit many others and accelerate...

... incredibly important to the stability of the biological research enterprise going forward, in this post...Addgene), that need people who are data oriented, scientifically minded, and able to communicate complex information...underrepresented groups under their mentorship with the same critical eye they place on others within their labs - as...

...Using AAV vectors in complex biological systems can be tricky at best, and downright infuriating at worst...promoter in vivo. Molecular Therapy - Methods & Clinical Development, 3, 16062. https://doi.org/10.1038... Expression specificity of GFAP transgenes. Neurochemical Research, 29(11 SPEC. ISS.), 2075–2093. https...

...sequences. Non-editing CRISPR applications direct catalytically dead dCas9 fused to a variety of epigenetic ...from Addgene. For most of these constructs, catalytically dead modifiers are also available as controls...Resources on the Addgene Blog Cas9 Activators: A Practical Guide CRISPR Between the Genes: How to Experiment...

...transposition: Class I TEs, also called retrotransposons, and Class II TEs, also called DNA transposons. DNA transposons...in a genome. But there is a type of mobile DNA, called a transposon, that’s a bit hyperactive and likes...

... a laughter program significantly decreased immunological markers of stress in adolescents. It also put...great use of your time even as that lab bench is calling, let me share my top five new favorite science ...

...understanding of how protein folding works and creating biological tools not found in nature. Dr. Baker’s team was...was not based on any existing protein structures, called Top7. Figure 1: Top7 receives the Nobel ...

...of an “electricity aggregation program” for all local households and businesses. Electricity aggregation... energy expenditure here at Addgene. These are critical systems that cannot be removed or substantially...

...preparations of several variants of a new optogenetic tool called PdCO, deposited by the Yizhar lab. Addgene is now...preparations of several variants of a new optogenetic tool called PdCO.  This is an "optoGPCR" (coupled to Go, so...

...with most reagents sent, and the most recent full calendar year When the website is launched, it will show...Amsterdam). He develops, characterizes, and uses genetically encoded fluorescent probes. You can follow him...

...September 13, 2018 This massive scale reagent sharing led us to calculate how much time we have saved researchers...

...wear a mask,↔️social distance, 💓show gratitude,☎️ call up an old friend. Remind them you care!#thanks #...viruses. Final tree with the complete #MicrobeAdventCalendar ornament set (plus more!) It features archaea...

...E6 regulatory element, which targets it to VIP cortical interneurons. Take a look to see if these new ...Wilson New serotype AAVrg = AAV retrograde (also called AAV2-retro) Looking for more viral vectors? Browse...

...:26 : Kwabena's plans for work in Ghana - Using local biomass "waste" to produce biofuels. 23:27 - 25:...foods. 44:15 - 46:34 : Kwabena's goal to build biological science opportunities in Ghana. 46:35 - end :...

...finishing up their time in the PhD Program in Biological and Biomedical Sciences at Harvard University. Ben and...

...acquires through Addgene to produce molecular biological tools like Taq polymerase, T7 polymerase, EcoRI...scientists to continue to do excellent work in the biological sciences. Future awardees Now that this first...

...the antigen of interest. The immortalized cells, called hybridomas, are individually isolated and expanded...repositories provide standardized quality-control and technical support to the community. Thanks to Our Guest ...

...Hiroshima University. “However, there had been no systematically established way of making an all-in-one vector...Activity Lentiviral CRISPR Libraries Enable Genome-Scale, Knockout Screening ...

...plasmids available at Addgene. We will still periodically focus on specific CRISPR plasmid tools more ...E. coli, the Chong Zhang lab developed a genome-scale CRISPR inhibition pooled library (~60,000 gRNAs)...

...Cynthia Fuhrmann at University of Massachusetts Medical School. It has been supported by a Steering Committee...A grant from the National Institute of General Medical Sciences (NIGMS) will support the development and...

... Support Specialists here at Addgene. If you’ve called our customer service line or sent an email to help...plasmids, store your plasmids or viral items, technical support, and more. ...