... Cell 18, no. 5 (2016): 573-86. PubMed PMID: 27152442. PubMed Central PMCID: 4871596.  5. González, Federico...phosphatase treatment, substituting cytidine with 5-methylcytidine, and/or substituting uridine with pseudouridine...by the piggyBac transposon." Nature Methods 6, no. 5 (2009): 363-69. PubMed PMID: 19337237. PubMed Central...PubMed PMID: 20952534. PubMed Central PMCID: 2956203 5. Robinton, Daisy A., and George Q. Daley. "The promise...methods do you have experience using? Let us know about your iPSCs experiences in the comments below! References...Additional Resources on the Addgene Blog Learn about Methods to Reprogram Peripheral Blood Cells Check...

...This is the third in a 5 part series on Management for scientists. Feedback is help and it should result...build it into your routines. This is the third in a 5 part series on Management for Scientists. Subscribe...employee about what they do well and where they need to improve, and they’re also clear about how the ...enough to tell them about areas they need to improve in. If a manager has concerns about an employee and ...language to talk about real performance issues. Watch this little video to learn more about indirect language...Medical School researcher Connie Cepko to learn about her management style. Formalizing the feedback session...informal feedback is a very effective way to bring about change in behaviors and habits in a nonthreatening...

...Bleomycin glycopeptide Induces DNA breaks Bactericidal 5-100 ug/mL Erythromycin macrolide Blocks 50S ribosomal...subunit; inhibits peptidyl translocation Bacteriostatic 5-25 ug/mL in EtOH *In prokaryotes.     **Dissolve...molecular biology, yet scientists rarely stop to think about how much easier it makes our lives. Plasmid transformation... sterile filter unless otherwise specified. The above table lists some antibiotics commonly found in the...or light – do not add them to media hotter than about 55oC and store plates/stocks wrapped in foil if ...

...outperform Cas9 in GC-rich regions. DNA guides NgAgo uses 5’ phosphorylated DNA guides (so called “gDNAs”) rather...if NgAgo preferred certain guide sequences - using 5 guides/gene for 10 genes, they did not observe differences...impaired NgAgo activity, with three mismatches abolishing activity. One-guide faithful NgAgo guide loading... how it’s working in the comments section! Read About NgAgo User's First Impressions Here References ... Additional Resources on the Addgene Blog Read About Cas9 Homolog Cpf1 Check Out High Fidelty Cas9 Mutants...

...consistent 5% positive error across the plate is fine; a 5% positive error on one side and a 5% negative...  Probe should have a melting temperature 5-10 degrees above the primer and be at least 25 bp away from... Probe is on a hairpin structure included in the 5’ primer Yes Yes. Agarose gel  LUX fluorogenic...

...Several behavioral assays can be done on zebrafish at 5 days post fertilization, including escape behavior...behaviors, the researchers measured the tail movements of 5 day old fish. The larvae were immobilized in agar ... posts about viral vectors Subscribe to the Addgene blog Resources on Addgene.org Learn about our ready-to-use...

... This is the first in a 5 part series on Management for scientists.  “I'm slowly becoming a convert ...carrot, and not the stick.     This is the first in a 5 part series on Introduction to Managing People for...the same as not being a bad manager (see cartoon above – what type of bad manager have you had?). You can...Managing scientists I should add a few special words about managing scientists because, let’s face it, we are...

...a free platform for sharing short, 3 to 5 minute videos about research. Videos can be shared by research... for communicating research. Short videos of 3 to 5 minutes provide more than enough time for researchers...measure the impact of their contribution. It offers a 5-step process and several examples for creating effective...learn about how researchers are using “big data” to look at Lyme Disease in new ways. Or learn about the...ot­her disciplines, are even less likely to read about our work. We therefore must discover new ways to.... Only on occasion do we then have time to read about research from other disciplines, or from other countries...communication through video     Video abstracts about research, especially published research, have grown...

...   The Mello lab also reported that modifying the 5’ modification of oligos used to generate linear dsDNA... animal.  PAM sequence considerations For changes 5’ to the PAM, sequences corresponding to the same strand... using two Cas9 targets. An orientation where the 5’ ends of the PAMs are facing outwards, ensuring the...Edraki A, Watts JK, Sontheimer EJ, Mello CC (2018) 5′ Modifications Improve Potency and Efficacy of DNA...learn more about genome editing with CRISPRs? Check-out Addgene's CRISPR Resources Learn about gRNA design...interest, I encountered low efficiency (0.13%) and laborious handling (screening 768 F1 animals). Recovering...

...of culture Approximate DNA yield** Miniprep 1–5 mL 5–50 µg Midiprep 25–100 mL 20–200 µg Maxiprep...for example. These kits are designed to handle ~1–5 mL of liquid culture. Using a higher volume can affect...µg Megaprep 0.5–2.5 L 1.5–2.5 mg Gigaprep 2.5–5 L 7.5–10 mg *Can vary from kit to kit, check the...columns we’ve been talking about!) are a significant source of waste in laboratories. To help combat this, ...dependent on many different factors as we detailed above, so be sure to pick what kit works best for you!...

...Cellular Biology 5(11): 3274–79. PubMed PMID: 2427927. PubMed Central PMCID: PMC369144. 5. Kranz, Andrea...Cellular Level in the Mouse.” Nature Biotechnology 21(5): 562–65. PubMed PMID: 12665802. 9. Villa, Katherine... on the Addgene Blog Learn More about FLEX-vectors Learn More about Cre-Lox Check out Retrograde AAVs...expression have been developed. Read on to learn more about FLP-FRT, Cre-lox, and how combinations of FLP and...

...their Roller phenotype. If SEC is inserted at the 5’ end of a gene, the SEC separates the targeted gene...are then heat shocked to remove the SEC and after 5-6 days of growth, wild-type animals that have undergone...just one injection step! The entire protocol takes about 2-3 weeks but requires less than eight hours worth... this species, namely, the robust HDR mentioned above, an accessible germline, and multiple selectable...modular system that requires very little hands-on labor and could facilitate the tagging of every protein...

...similarly but not identically to BE3, modifying a 3-5 base window 18 bases upstream of the PAM.  David Liu...than BE2 Nishida et al., 2016 Target-AID   Edits 3-5 base window surrounding -18 position upstream of the...CRISPR-based cytosine base editing. Nature Biotechnology, 41(5), 663–672. https://doi.org/10.1038/s41587-022-01532...generated from TadA variants. Nature Biotechnology, 41(5), 686–697. https://doi.org/10.1038/s41587-022-01611...low off-target activity. Nature Biotechnology, 41(5), 673–685. https://doi.org/10.1038/s41587-022-01533...Homology-Directed Repair Learn about base editing for plants Read about four base editor reporters Additional... The BE3 system increased editing frequency to above 30% for a variety of targets in human cell lines...

...the protein with enterokinase, which recognizes the 5 amino acids on the C terminal of the tag. Of the most... most common tags, FLAG is the most charged, with 5 negatively charged and 2 positively charged amino ...Tags have a lot of major benefits – as discussed above – but there are a few instances when they should... hindrances, and destabilization can be brought about by the tag so it’s important to validate tagged ...

...AAV9, AAVrg pAAV-hSyn-DIO-mCherry 37825 AAV1, 2, 5, 8, 9 (20 µL size serotype testing) pAAV-CAG-GFP ...size serotype testing) pAAV-CAG-GFP 50465 AAV1, 2, 5, rg (20 µL size serotype testing) pAAV-hSyn-EGFP ...vector name and serotype you’d like to be notified about. Controls Plasmid Serotype Name 37825  AAVrg...vectors 101 eBook Resources on Addgene.org Learn about Addgene's viral service Find viral protocols and...

...expected to be >12 kb. However if it recombines at the 5’ and 3’ LTR regions, it splits into two smaller plasmids...recombined vector backbone.   Ready, set, grow! Our 5 Tips for Troubleshooting Viral Transductions will ...molecular biology and viral vector protocols Learn about the common lab E. coli strains Find tips for viral...

... codons/second. They also explored the effects of 5’ UTR variants in translation of cell cycle protein...Emi1, showing that ~80% of transcripts with a long 5’ UTR were translationally silent. Both of these methods...Additional Resources the Addgene Blog Learn More about SunTag Catch up on Your GFP Background Choose the...

...Coli Synthetic Biology.” ACS Synthetic Biology, vol. 5, no. 10, American Chemical Society (ACS), 2 May 2016...Traditional Biology.” ACS Synthetic Biology, vol. 5, no. 1, American Chemical Society (ACS), 4 Nov. 2015....” PLOS ONE, edited by Alvaro Galli, vol. 13, no. 5, Public Library of Science (PLoS), 30 May 2018, p.... vol. 8, no. 11, American Chemical Society (ACS), 5 Nov. 2019, pp. 2593–606. Crossref, https://doi.org... constructs that compose genetic circuits and metabolic pathways. For many MoClo systems, the individual...interested in recombinant protein purification and metabolic engineering. If instead you want to build genetic...designed for flexibility. The Fragmid kit described above works in both mammalian systems and Drosophila. ...

...the success of precise change via HDR limited to 0-5%. This blog post covers a brief review of different...SpCas9 rAPOBEC1 NGG 6 SpCas9 Target-AID (PmCDA1) NGG 5 SpCas9-VQR rAPOBEC1 NGA 8 SaCas9-KKH rAPOBEC1 ...PMID: 25733849. PubMed Central PMCID: PMC4371917. 5. Shimatani, Zenpei, et al. "Targeted base editing ...gene, Xw, except for a single base. If you heard about CRISPR, one question probably comes to mind: can...compatibility All of the base editors mentioned above are SpCas9 nickase-based and depend on the presence... HDR methods for rice crop improvement in the laboratory of Yinong Yang at PSU.   References 1. Komor,...The CRISPR-Cas Revolution, Cold Spring Harbor Laboratory (2018). Additional resources on the Addgene blog...

...methodology with T cell receptor engineering (Ref 5)  as described by Blue Flame Addgene depositor Professor...PMID: 26343581. PubMed Central PMCID: PMC4597191. 5. Eyquem, Justin, et al. "Targeting a CAR to the TRAC...Cancer, Inflammation and Immunity and learn more about “... recent findings in basic immunology, cancer-immune...curing cancer in cross-disciplinary projects and collaborations. Summarized by Professor Michael Karin from...This was followed by a talk from Professor Karin about alcohol and obesity as driving factors for chronic...got a chance to talk with Dr. Friedmann-Morvinski about tools used to analyse the tumor microenvironment...community, contact [email protected] to find out more about our free deposit and distribution service! References...