We narrowed to 847 results for: CAL;
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TypeBlog PostPublishedFeb. 11, 2020, 2:15 p.m....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...
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The Importance of a Fun Workplace: Company Culture at Addgene
TypeBlog PostPublishedSept. 3, 2015, 2:30 p.m.... 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 ... -
Negotiating Work and Life: How to Find the Joy
TypeBlog PostPublishedMarch 20, 2014, 3:04 p.m....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,... -
How to Write a Scientific Review Article
TypeBlog PostPublishedFeb. 16, 2017, 3:30 p.m.... 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... -
Save Time with Transient Plant Leaf Transformations
TypeBlog PostPublishedJuly 25, 2019, 12:03 p.m....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 ... -
Overcoming the AAV Size Limitation for CRISPR Delivery
TypeBlog PostPublishedSept. 16, 2020, 1 p.m....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 ... -
The PAM Requirement and Expanding CRISPR Beyond SpCas9
TypeBlog PostPublishedAug. 20, 2020, 2:30 p.m.... 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... -
Four Base Editing Reporters to Monitor and Enrich Editing in Real-time
TypeBlog PostPublishedJuly 7, 2020, 1:15 p.m....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. ... -
Tips for Writing a Good Cover Letter
TypeBlog PostPublishedMay 21, 2024, 1:30 p.m....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... -
CRISPR 101: Which Cas9 Do I Choose for My CRISPR Experiment?
TypeBlog PostPublishedOct. 13, 2022, 1:45 p.m....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... -
Antibodies 101: Reading a Flow Plot
TypeBlog PostPublishedFeb. 6, 2024, 3:13 p.m....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 ... -
Viral Production at Addgene
TypeBlog PostPublishedOct. 24, 2023, 1:15 p.m.... 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... -
Binning Singletons: Tackling Conference Networking When You Don’t Know Anyone
TypeBlog PostPublishedFeb. 18, 2020, 2:15 p.m....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... -
Four Ways to Package Transgenes That Exceed the Size Limit of Adeno-associated Virus
TypeBlog PostPublishedSept. 1, 2020, 1:15 p.m....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... -
CRISPR 101: Validating Your Genome Edit
TypeBlog PostPublishedNov. 3, 2022, 12:15 p.m....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... -
Transferable Skills Guide: Time Management
TypeBlog PostPublishedDec. 5, 2017, 1:52 p.m....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... -
CRISPR 101: Engineering the Plant Genome Using CRISPR/Cas9
TypeBlog PostPublishedOct. 11, 2016, 2:30 p.m....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 ... -
Site Directed Mutagenesis by PCR
TypeBlog PostPublishedAug. 2, 2016, 2:30 p.m....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... -
Targeting HIV-1 with CRISPR: Shock and Kill or Cut it Out?
TypeBlog PostPublishedMay 10, 2016, 2:30 p.m....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... -
Thoughts on the Future of Research 2015 Boston Symposium
TypeBlog PostPublishedNov. 25, 2015, 4 p.m.... 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...