...transferred between cells. At the time, these extranuclear agents of heredity were thought of as everything...first plasmid “cloning” experiment. Dr. Cohen and colleagues treated a tetracycline resistant plasmid, pSC101...topology, and cloning and gene expression. Just released: Addgene's Plasmids 101 (2nd Ed.) eBook And now...and more pictures and pro tips.  Happy reading! Learn more about the history of plasmids and cloning: ...

...few quirks specific to the PCR cloning process - please check out that post if you need a more detailed...PCR primers for molecular cloning consist of: Leader Sequence: Extra base pairs on the 5' end of the... existing software (a quick internet search will lead you to here and many others). If we put the sequence...product. Additional Resources on the Addgene Blog Learn About Other Cloning Techniques Catch Up on All Things...

...transduction.) Compared to CCP transduction, iTOP is at least four times more efficient in primary fibroblasts... gRNA, which subsequently enter the nucleus and cleave the specified target. Figure adapted from D’Astolfo...expression is by nature transient, which may lower nuclease toxicity or the potential for off-target effects... Biol. 2000 Jul;10(7):290-5. PubMed. Is VP22 nuclear homing an artifact? Lundberg M, Johansson M. Nat...

...indicated that a reasonable rule is to require at least 96% sequence identity when detecting a reference...commercial plasmids were inconsistent and sometimes clearly wrong. In an effort to be rigorous, we dug into... harness common feature detection in a new way, please contact us at snapgene.com. Many thanks to our...Blog Read about our SnapGene powered plasmid maps Learn how we use SnapGene during the plasmid quality control...

...interesting at an event. Or, perhaps you have a colleague who seems fun and helpful who you’d like to get...connections. I mention this because I want it to be clear that it is completely impossible for me or anyone...contacts over the years in many different ways. This leads me to my last tip: It’s usually not worth connecting...networking Attending a conference soon? Get tips here Learn about informational interviews ...

...night of the iPSC symposium, a panel of thought leaders in the field--including Shinya Yamanaka, George... of ethics can be daunting because they have no clear cut answers, I applaud the stem cell community for...Disease; and Kevin Eggan presented work that is leading to promising clinical trials for ALS.   I have ...that!  Additional Resources on the Addgene Blog Learn about Efforts to Fight Rare disease with iPSCs Read...

...it unuseable. With the large size of the SpCas9 nuclease (~4.2 kb) this is a particularly prominent problem... perform an optimization-scale infection and at least one screening-scale experiment achieving the gRNA...the publications associated with these libraries. Please note, the pooled libraries we distribute in lentivirus...or are thinking about using our viral services. Please send any questions or comments to help@addgene....

...viral vectors. Here are some of the AAV we have released in the last few months. You can also browse our...coming soon! These vectors listed below should be released at Addgene in the next 6 months, pending quality... particular catalog item is available for order please email [email protected] with the vector name and...viral vectors 101 eBook Resources on Addgene.org Learn about Addgene's viral service Find viral protocols...

...of happen with Cas13. CRISPR-Csm degrades both nuclear and cytoplasmic mRNA with 90-99% knockdown efficiency...CRISPR-Csm presents an attractive alternative for nuclear-RNA targeting, where siRNAs are ineffective and...expression changes of non-target genes and RNA-seq of a nuclear (MALAT1) and cytoplasmic (CKB1) RNA target. Adapted...antibody per target just isn’t enough and we recently released Anti-GFP [N86/8R] as another option for targeting...

...Addgene blog Learn about other CRISPR methods for bacterial genome engineering   Learn more about lambda... novel genetic variants in bacteria. Read on to learn about the key components of EvolvR and its potential...extends it from the 3’ end, while its native endonuclease activity degrades the displaced strand.   The...

...embryo—even if it’s not necessarily via CRISPR/Cas9. This leaves us with the second critique. Absent of a broad ...intended parents is “rehoused” in a separate, enucleated zygote with healthy mitochondria, with the intent...Nuffield Council’s report is that when it became clear that this assistive reproductive technique was feasible...featuring a clinical pregnancy, was rejected by at least one academic journal for exactly the critiques leveled...

...accessible to everyone but these projects could also lead to novel yeast creations that may not be pursued... the different samples. The goal is to not only learn something about the microbial diversity in different... to find a community lab in a city near you. To learn more about what we’re up to or how you can help ...Disease & Open Science: Interview with Darrell Kotton Learn How Plasmid Sharing Speeds Science Resources at...

... work to enable future discoveries. Read on to learn more about Xin and Fatima's work and keep checking... work at the 2017 Winter qBio Meeting. You can learn more about Xin’s work with photoswitchable components...enzyme breaks down the HMO of interest, the lactose released activates expression of GFP from the lac operon...Biology! Additional Resources on the Addgene Blog Learn More about Photosactivatable Fluorescent Proteins...

...stakeholders to learn more about their needs and any problems that they are trying to solve. After learning more ...Technologies, I work with our customers in the US to learn where the gaps are in their sequencing workflows...problem-solver. A lot of the time, the problem to solve is clear, but the solution is more complicated. You’ll need...

...experiments. As a common example, a selection will leave you with the colonies that contain your plasmid ...negative controls for your PCR.  Find the details and learn more about colony PCR in this blog post.   ...can confirm the sequence of the entire plasmid. Please note that NGS verification is more time-intensive...biology protocols and tips Resources on Addgene.org Learn more about Sanger sequencing Read the Molecular ...

...are some of the AAV and lentiviral preps we have released in the last few months: Lentiviral preps for ...coming soon! These vectors listed below should be released at Addgene in the next 6 months, pending quality... particular catalog item is available for order please email [email protected] with the vector name and...viral vectors 101 eBook Resources on Addgene.org Learn about Addgene's viral service Find viral protocols...

...fluorescent reporter assays to quantify targeted Cas9 cleavage, and repair through homology directed repair (...while cells containing the reporter gene that was cleaved by Cas9 and repaired via HDR using the supplied... seamlessly remove the integrated GFP reporter leaving you with just your desired genomic edit.  Find ...using GEmCherry2. In this experiment they saw a clear difference in mCherry expression using FACs sorting...

...collection in your experiments and published your data, please fill out this form and we’ll add your citation ...combinations of WT, codon optimized, untagged, cleavage sites (thrombin and TEV), and C-terminal tags ...downstream experiments and offer flexibility in tag, cleavage, codon usage, and expression system. Mammalian...Addgene blog Read all of our COVID-19 blog posts Learn about other COVID-19 plasmids available at Addgene...

...Additional Resources on the Addgene Blog Learn More About Lentivirus Learn more About AAV Get the Scoop on Our...of 2016. The top viruses of 2016 were (drumroll please)... Top Lentivirus: lentiCas9-Blast Most of Addgene... to perform unique editing tasks, classic Cas9 nuclease, which introduces double stranded DNA breaks, ...

...the nucleus through nuclear pores and thus do not require breakdown of the nuclear envelope during cell...there – most of which we would all like to steer clear of! However, a subset of viruses are commonly used...which ones could be helpful to you. Interested in learning more about a specific virus? We have you covered...