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...of the hood while work is in progress.Learn more about cell culture contamination in this post from the... bath constantly at 37°C for this purpose.Learn about the effects of CO2 on sample pH Cell morphology:... Set up a routine to split cells Don’t be lazy about splitting cells, instead try to form a routine. ...secretion of lactic acid (a by-product of cellular metabolism that is toxic to the cells), and need to be split...

...specific genes. Many years ago, people in the laboratory became frustrated with using glass cloning rings...the hood. A 48-well plate is also prepared with about 200 ul of culture medium containing serum in each...incubator, additional growth medium is added to about halfway up each well. Tips for using the agarose...with the firmer 2% agarose gel solution discussed above for picking. Many thanks to our guest blogger Iris...

...the context of the cell. However, much knowledge about functionality can be gained by understanding the...subcellular marker proteins, it’s possible to get an idea about the subcellular structures your protein resides ...with. These “colocalization” studies give insights about the proximity of two proteins within the same subcellular...Additional Resources on the Addgene Blog Learn More about Plasmids for Endogenous Gene Tagging in Human Cells...

...technology. To dig more into what researchers felt about NgAgo, I conducted a survey and asked researchers...successful, and the second section asked basic questions about the NgAgo protocols used by respondents (transfection...27136078. 2. Burgess, Shawn., et al. "Questions about NgAgo." Proteins & Cell (2016). 3. "NgAgo: Genome-editing...PMC4343198. Resources on the Addgene Blog Learn More about NgAgo Enhance CRISPR Targeting Specificity with ...

...directly back to your specific goal. When thinking about step 3, however, you may be surprised at the number...Delivery methods for mammalian cell lines As mentioned above, the expression system you choose in many ways dictates.... The user should review the current literature about their preferred model.  If your expression system... Check out our CRISPR Featured Topic Page Learn about CRISPR Delivery Using Ribonucleoproteins (RNPs) ...

...genes above with your gene or genes of interest. Some of the plasmids listed in the table above (and their... an internal region of the mRNA. In the figure above, you can see that the IRES element acts as another... first discovered in picornaviruses, are short (about 20 amino acids) and produce equimolar levels of ...

...CRISPR Featured Topic Page Learn about Non-Homologous End Joining Learn about Base Editing Read 3 Tips to ... post on increasing HDR efficiency to read more about optimizing this distance. Homology arms and types...screening strategies for your experiment. Also learn about Addgene’s knock-in plasmids for reagents to help...

...Read this blog post about CRISPR Kinome Libraries available at Addgene Learn about the advantages of pooled...large-scale functional screens Read this blog post about genome-wide screening using CRISPR Resources on...libraries for the mouse genome! Other Resources Read about CRISPRi and CRISPRa from the NIH E-newsletter ...

...gene therapy applications. Read on to learn more about the current (and future) applications of lentiviral...do both, Addgene depositor Didier Trono and collaborators turned to the lentivirus HIV-1, well-known to...Libraries Enable Genome-Scale, Knockout Screening Read about Genome-Wide Screening Using CRISPR/Cas9 Additional...Addgene.org Check out Our Lentivirus Guide Learn about Virus Biosafety Find Lentiviral Vectors for Your...

...genome editing than Cas9 - read on to learn more about Cpf1 and check out our CRISPR guide for a refresher...mediated cleavage are 42-44 nucleotides in length, about the same size as Cas9’s, but with the direct repeat... Cpf1 was first identified, we didn't know much about its on-target and off-target editing efficiency....CRISPR system proved we had a lot more to learn about CRISPR biology. Later work, like the adaptation ...

...on the Society for Laboratory Automation and Screening (SLAS) Electronic Laboratory Neighborhood.   Work...worry less about how they will handle an emergency and this will make you much more confident about their ...

...all, a delivery method. I would be remiss to talk about them without looking at what they’re delivering .... But in the years since, we’ve learned so much about what it takes to make these viral vectors not only...., 2016), video, educational resources, and collaborations, and will continue to do so, in the hopes that...leave a comment; or take our AAV survey. Learn more about GCaMPs and other tools developed through the GENIE...

...Resources on the Addgene Blog  Read more about Cpf1 Read more about SaCas9 and AAV Check out our CRISPR Featured... and VRER) recognize novel PAM sequences (shown above). The VQR, EQR, and VRER Cas9 variants are capable...example, Cas9 from Staphylococcus aureus (SaCas9) is about 1 kilobase smaller than SpCas9, so it can be packaged...

...Addgene Blog Learn about using toehold switches to detect Zika virus   Learn more about targeting RNA with...time-consuming, and complex to use outside of standard laboratories. In the case of the COVID-19 pandemic, qPCR ...with Cas13a (C2c2) Learn more about Cas12a (Cpf1) and its multiplex genome editing abilities Resources...

... an AAV rookie  Learn about some considerations when using AAVs Learn more about using AAV as a tool for...PubMed PMID: 29175632. If you want to learn more about how PHP.B and PHP.eB were engineered, check out ...Additional resources on the Addgene blog Learn more about PHP.eB, and PHP.S, a peripheral nervous system (...

... to target individual genes can be relatively laborious because multiple plasmids/viruses are needed to...screening strategies Although the tools described above make CRISPRa more accessible to neuroscientists,...vitro and in vivo applications, the tools described above make CRISPRa in the brain relatively easy and accessible... CNS using CRISPRa as a member of the Gelbard Laboratory at URMC.     References Liu, Yanxia, et al. "...

...RSpace is integrated with most of the tools noted above, and has a modern API. Integrations with additional...into RSpace. The kind of informal conversations about research that can take place in Slack normally never...RSpace documents to external data sources (described above) further enhances the value the repository integrations...Minisymposium on Reproducibility Read all blog posts about scientific sharing Resources on Addgene.org Find...

...that lead to these preferences and what can we do about them? Read on to find out!   Why do organisms have...understood, but some possible reasons include: Metabolic pressures - it takes cellular resources to produce...amino acid I’d like to encode,” but, as described above, not every region of a protein should necessarily...Effects on transcription (remember, it’s not all about translation - e.g. codon choice may interrupt transcription...

...resistant infection or not, because scientists in the laboratory will need to isolate and grow whatever bacteria...method that is frequently used in the clinical laboratory. What can we do to stop the emergence of antibiotic...CDC. Antibiotic resistance can be scary to think about, but we can all do our part to learn all we can,...drug resistance. Additionally, she is passionate about health equity, science communication, and public...

... organisms (Chavez et al., 2016). To learn more about the different types of Cas9 activators, check out...covering VP64, Suntag, SAM, and VPR. Worry less about off targets Unlike Cas9 cutting activity, off target...of the Addgene Scientific Advisory Board. His laboratory at Columbia University is focused advancing scientific... Variant for Your Next Experiment Find out More about Genome-wide Screens with CRISPR/Cas9 Learn How CRISPR...