...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...

... No Roth 83896 pAAV-hDlx-GiDREADD-dTomato-Fishell-5 hM4D (Gi) hDlx tdTomato No Fishell 83897 pAAV-hDlx-GqDREADD-dTomato-Fishell...Learn more about chemogenetic resources available at Addgene. See our inventory of chemogenetic constructs...your experiment. For a more complete description about chemogenetic receptors, ligands used for chemogenetic...

...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. ...

...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...

...phosphoserine directly into recombinant proteins (5). Optimization of this system defined three components...into recombinant proteins, such as the kinase MEK1 (5). MEK1 is inactive without requisite dual phosphorylation...307.5717 (2005): 1969-1972. PubMed PMID: 15790858. 5. Park, Hee-Sung, et al. "Expanding the genetic code... one of the MEK1 phosphomimetic residues in the above study with phosphoserine (to create a pSer/phosphomimetic...non-phospho species on PhosTag gels as described above. Reagents and protocols Sep-OTSλ plasmid - Addgene...interested in the intersection of cell signaling, metabolism, and the mighty mitochondrion. You can follow...Blog Verify Your Constructs with Colony PCR Read about Tags for Protein Purification Resources on Addgene.org...

...provides both the endonuclease and ligase function while 5’ adenosine overhangs left by Taq perfectly anneal ...articles; so if you are interested in learning more about any one of them, please click on the links throughout...laying around the lab. If not, you’ll be looking at about $50-$60 per enzyme. Once you’ve got a few of these... relatively inexpensive and can be obtained for about $70, resulting in a price of a few dollars per reaction...

...downstream of the promoter, you can add an EcoRI site just 5’ of the start of the gene and a HindIII site just ...two HindIII digested ends will anneal leaving the 5’ end of your gene just downstream of the promoter ...plasmid : insert ratio” in the ligation reaction. 5. Purify the finished plasmid   Once it looks like... (post or pre run in the table above, respectively). Some of the above stains require you to visualize...equivalent to backbone + insert, unsuccessful 2 and 3 above), you can try ligating the recipient plasmid alone...get experimenting!   Additional Resources: Read about Gel Stains on the BitesizeBio Blog  Additional ...

...makes it so forgiving, is the incorporation of a 3’-5’ exonuclease in the reaction mixture. The exonuclease...expression cassettes for yeast transformation can take 5 days. NEBuilder shortens that time to one day by simplifying...scarless plasmid inserts.  Want to Learn More about DNA Assembly? Check Out Our Plasmids 101 eBook! ...to produce sgRNA for CRISPR experiments can be laborious, requiring multiple cloning steps. You can simplify...Additional Resources on the Addgene Blog Learn More about Gateway Cloning Gibson Assembly and Other Long-Homology...

...restriction enzymes that cut DNA at specific sites (5). Imagine that you have a genome with unknown sequence... allowed for many seminal scientific discoveries (5, 8). While newer techniques have largely replaced ...PMID: 19283728. PubMed Central PMCID: PMC2743315. 5. Roberts, Richard J. "How restriction enzymes became...abundant in a sample. Whereas Southern blotting is labor intensive and requires a large amount of high-quality...DNA technologies that are now commonplace in laboratories. As scientific technologies rapidly progress...PMC4411503. Resources on the Addgene Blog Learn More about Restriction Cloning Get Tips on Analyzing DNA Sequencing...

...recent paper to see the Gal4/UAS system in action (5). I use the Gal4/UAS system to knock down gene expression...Drosophila, 3, 1–10 (2015). PubMed PMID: 21356876. 5. J. Chow, Z. Marka, I. Bartos, S. Marka, J. C. Kagan... use the Gal4/UAS system, and you can read more about them in review articles (2–4). You can also take...

...Zhang et al. designed multiple gRNAs to target the 5’ long terminal repeat (LTR) of the HIV-1 genome, which...Kaminski et al. designed gRNAs to target the HIV-1 5’ and 3’ LTRs and expressed them along with Cas9 in...Replication and Accelerate Viral Escape. Cell Rep. 2016 Apr 5. PubMed PMID: 27068471. Ramanan V, Shlomai A, Cox ...reservoirs, chronic inflammation, and negative metabolic effects of ART. Why can’t the body mount an immune...

...protein, activating its membrane fusion function.( 5 ) TACE - (ADAM17) a metalloprotease that cleaves hACE2...lightning speed to elucidate as much as they can about this novel virus. It is now known that SARS-CoV-...to distribute SARS-CoV-2 plasmids. To read more about this partnership and the plasmids that are available...and open-access SARS-CoV-2 detection assay for laboratory and home testing. Kellner, et al. bioRxiv 2020.06.23.166397...

...Advice on Picking a Graduate Lab and Project Check Out 5 Science Videos from Tom McFadden Resources at Addgene.org...of our Hot Plasmids podcast series, you'll learn about optogenetics tools for controlling protein activity...

...Advice on Picking a Graduate Lab and Project Check Out 5 Science Videos from Tom McFadden Resources at Addgene.org...of our Hot Plasmids podcast series, you'll learn about new red fluorescent proteins, AAV tools for targeting...

...Addgene Blog 5 factors to help you choose the right cloning method Find blog posts about scientific sharing...scientists save even more!) If you assume an average of 5 hours of hands-on time per plasmid, Addgene, since...cloning before you can start your experiment? What about all the reagents you need to order? Sometimes, it...dive deep into generating the data you’re excited about. When we were closing in on the one million plasmids...million plasmids shared than not!), we think it’s about time we updated this post. How much time does ...

...LTRs define the limit of what will be packaged. The 5’ LTR and 3’ LTR, respectively, act as RNA polymerase...adenovirus types, but most vectors are based on adenovirus 5 (Ad5).     Fig. 3 - Viral vector components...aren’t reflected in the common components outlined above for gamma-retroviruses. While not required for lentiviral...and helper plasmids for all the viruses discussed above (gamma-retrovirus, lentivirus, adenovirus, and adeno-associated.... Cold Spring Harbor (NY): Cold Spring Harbor Laboratory Press; 1997. Principles of Retroviral Vector ...

... Antibody Validation proposed 5 pillars for antibody validation.  The 5 pillars include genetic strategies...Narrowing the field After working your way through the above steps, you may find that you still have a number...advice if you need to troubleshoot. What do you know about the vendor and the vendor’s technical support? An...validation is equal and you should think critically about what data is and is not shown on an antibody website...

...set at .05, then 5% of the time the statistical test will be incorrect. If you have 5 samples and perform...comparison is needed, without a high number of samples (~5). Tukey’s test – Most conservative test. Higher false...a summary snapshot without telling you anything about outliers or data spread within those data sets. ...the rules” and which we have certain information about. The assumptions and requirements that need to be...Quantitative If your data doesn’t fit the requirements above, a non-parametric test may be the way to go, which...only performing the comparisons you really care about. Family-wise error is the probability of a false...rate than the multiple comparison tests described above. Violated assumptions and statistics Statistical...