... to generate two more OptoNBs, one against mCherry and one against GFP. Insertion of AsLOV2 into loop ...improve this process. The team used nickel-coated agarose beads coated with his-tagged OptoNBs. OptoNB coated...

...luminescent imaging a reality, Addgene depositor Takeharu Nagai and colleagues at Osaka University have developed...signal. Since their first publication in 2012, the Nagai laboratory has assembled a collection of multicolored...imaging. Nano-lanterns and your experiments The Nagai laboratory has a done a wonderful job characterizing...

... preliminary injunction against the FLSA updates was granted nationwide. Again, we at Future of Research...This post discusses how a nationwide injunction against the FLSA is affecting universities' decisions to...

...: Forward oligo: 5’- CACCGNNNNNNNNNNNNNNNNNNNGTTTGGGTCTTCGAGAAGACCTATTCXXXXXXXXC -3’; Reverse oligo: 5... 5’- AATTGXXXXXXXXGAATAGGTCTTCTCGAAGACCCAAACNNNNNNNNNNNNNNNNNNNC -3’, where NNN and XXX represent the ...

...However, the cloning process — including the propagation of plasmids in bacteria — has many points at ...called Geneious. Here, the raw reads are mapped against a reference sequence, if one was provided. If no...plasmid, and the full QC process is started over again after receiving the replacement plasmid at Addgene...

...Please indicate which growth strain Addgene should propagate your plasmids in: DH5α, NEB Stable, ccdB Survival...Survival, Pir1, or other. Whenever possible, we propagate plasmids in the standard cloning strain DH5α. For...choose the Pir1 strain. If your plasmid cannot be propagated in any of these standard strains, choose other...

... transcription of gene III, permitting phage propagation and keeping that dCas9 sequence in the pool. ...with BioRender.com. After 24 days of phage propagation, Hu et al. isolated xCas9 1.0-1.4 and subjected...were capable of cleavage, they tested the clones against an NNN PAM library in bacteria where cleavage leads...

...spreads through the wild population and immunizes it against the first drive. Laboratories interested in building...question), why not use them whenever applicable?Against this, consider the cost of an accidental release...accidental release could delay real-world applications against scourges such as malaria and dengue for many years...

...experimental species. Discovering ways to culture, propagate, transform, store, and share all of these new ... eat a caterpillar once, vomit, and never do it again. Cichlids (Scott Juntti) Cichlids are 2,000 fish...cephalopods die after they lay their 1 clutch of eggs? Propagation is thus quite difficult. CRISPR genome editing...

...gut-wrenching, but you can get that feeling of camaraderie again in a non-academic career. You might not be in a ...you still find the idea of not holding a pipette again upsetting, there are ways to get back to the bench...opportunities. You can get your hands in nitrile gloves again! The opportunities are there once you start looking...

...tryptone, 5 g/L yeast extract, and 5 g/L NaCl) or on LB-agar plates containing glucose and L-arabinose at 37oC...allowing us to reduce the genetic footprint yet again. This also led to improved expression homogeneity...ultrasound imaging. If needed, resort the cells again for improved purity and contrast. For in vivo experiments...

...reprogramming efficiency through the expression of a shRNA against p53, a modified reprogramming factor cocktail, ... pCXLE-hOCT3/4-shp53-F 27077 OCT3/4 and shRNA against p53 pCXWB-EBNA1 37624 transient EBNA-1   ...19668191. PubMed Central PMCID: PMC2917235. 11. Nakagawa, M., et al., Promotion of direct reprogramming...

...lines expressing galactosyl(alpha1-3)galactosyl (alphaGal) sugars were less stable than viruses produced...express these sugars. (Takeuchi et al., 1997). The alphaGal sugars end up in the envelope and are targets ...targets for complement-based killing by anti-alphaGal antibodies. The VSV-G envelope protein is inactivated ...

...every experiment, an investigator needs a standard against which the results can be compared; results from...single experiment wherein Plasmid A (encoding shRNA against human Gene X in backbone Y) was transfected into...included as these provide the baseline/standard against which other samples can be compared. The Empty ...

...then revise their hypotheses and the cycle begins again. No part of this process is inherently political...gutting 38 EPA programs: draft budget  Science Magazine: A grim budget day for U.S. science: analysis ...analysis and reaction to Trump’s plan Science Magazine: NIH, DOE Office of Science face deep cuts in Trump’s ...

...secondary antibody that recognizes the primary, wash again, then image. But here’s where the “sequential” part...add the new primary, wash, add secondary, image again, inactivate, and repeat these cycles for all your...yourself. If you can’t find a suitable antibody against your target protein, consider fusing a protein ...

...abilities, we’re gaining new ground in the fight against cancer in its many forms. The dichotomous interactions...sequence of immune reactions that aid in the fight against cancer. Sipuleucel-T by the Dendreon Corporation...will more broadly activate the immune response against tumors. Some chemotherapeutic agents induce immunogenic...

...Distribution Please indicate which strain Addgene should propagate your plasmids in: DH5α, NEB Stable, or ccdB Survival...Survival.­ Whenever possible, we propagate plasmids in the standard cloning strain DH5α. Please indicate...indicate if your plasmid cannot be propagated in DH5α. For plasmids with highly repetitive sequences (which...

...approach, which focuses on validating antibodies against markers of the human blood leukocyte populations...on human blood leukocytes, including antibodies against these markers. The 371 CD markers characterized..., and include information on tested antibodies against the target. Helpfully, this also includes clone...

...scientists delivered some spectacular photos once again this year! All winners receive an Addgene care package...(@ChiTownBio) and one of our favorite twitter #agarart accounts, Petri dish Picasso (@PetriPicasso) came...

...winners of the 2019 contest. Agar art For the second year in a row, agar art is one of the winning decorations...

...ladders, the 100 bp ladder works well for both agarose and native polyacrylamide gels. Get Tips on Verifying... to Analyze Your Restriction Digests Make Your Agarose Gel Other Resources Read about Gel Stains on ...

...impressive science-themed holiday decorations yet again. This year, we had two different categories: one...his Christmas tree! It is that time of the year again: miniature #Christmas tree goes up at home!This ...

...Court’s decision to ban gene patents in a case against Myriad, an organization called Free the Data launched...of 304 open access publishers. By the time the magazine went to press, no less than 157 of those journals...

... antibodies drive an effective immune response against a pathogen. The Fc domain creates a link between...Inhibitory Fc receptors modulate in vivo cytotoxicity against tumor targets. Nat Med. 2000 Apr;6(4):443-6. doi...

...plasmid and the insert. By using the “Simulate agarose gel” option with the desktop version of Snapgene...more enzymes.   Click here to learn how to run an agarose gel     We’ve worked with Snapgene to update our...

... fraction of readers cover the same ground again and again as they don’t save the insights they glean ...

...Senior Director of Biology, is helping us fight against FOP in two ways. First, Addgene is distributing...plasmids with Addgene and join us in the fight against FOP. Second, Addgene will be hosting the AJ’s Boston...

...partial loss of an ITR during routine plasmid propagation, with complete ITR loss abolishing AAV packaging...packaging. To prevent ITR loss, it’s best to propagate AAV constructs in recombination-deficient bacterial...

...applications. But some techniques require you to select against the presence of a transgene instead, which can’...are currently used in these situations to select against CRISPR positive plants, but screening these can...

...THE RESULTS DISCUSSED BELOW Biologists are going gaga over the newest gene-editing protein - a DNA-cleaving...prokaryotes, and some of these proteins protect against invading DNA. In 2014, Swarts et al. characterized...

...final Forward Primer sequence of 5'-TAAGCAGAATTCATGTGGCATATCTCGAAGTAC-3'. For the Reverse Primer, the design...final Reverse Primer sequence of 5’-TGCTTAGCGGCCGCTCAGTACTTCGAGATATGCCA-3’. Preparing the PCR product for...

...lost for an age) fight in the Napoleonic Wars and against something even more sinister. 1000 pages long. ...reads, some are hard. Some I’ll never think about again, some I loved so much I’ll force them on everyone...

...mutations and truncations that can make vector propagation and sequencing a headache. How do I maintain ...integrity and prevent unwanted mutations?  During propagation, mutants rapidly replicate in a growth culture...

...F, Clouaire T, Leung JW, Cammarata M, Perez M, Agarwal P, Brodbelt JS, Legube G, Miller KM Genes Dev. ...small, sensitive immunohistochemical reagents. Yamagata M, Sanes JR Proc Natl Acad Sci U S A. 2018 Feb...

...experiment by simply running it unchanged over and over again, and the same goes for your workflow. Next time ...mistake, instead of silently vowing never to do that again (I’ve been there!), think of ways you could change...

...each one requires different elements for vector propagation. This post, along with a future companion post...yeast plasmid) allows for 50+ copies to stably propagate per cell. The copy number of these vectors can...

...achieve my purpose? Should I go over this slide again? Say, 'Oh, I forgot to mention X, so let's quickly... freezes and then crashes. You’re muted on Zoom again.  These things don’t just happen to you or me (though...

...for every assay. Antibodies that are generated against a linear antigen tend to work very well for applications...compare the staining patterns of multiple antibodies against the same target; tagged protein strategies that...

...His work focuses on the enzyme hydropersulfide-propagating enzymes 3-mercaptopyruvate sulfurtransferase ...mouse cortex. Image from He Tian. Congrats again to all the recipients! ...