...plasmids. Second generation packaging systems use 3 plasmids; an envelope plasmid that usually encodes...vessel size. Opting for a larger vessel, such as a 3 or 5-layer flask will require some initial optimization...available systems. Producing lentivirus: it's all about the T's (transfection and titers) To produce lentivirus... optimize transfection conditions. As mentioned above, plasmid ratios can be adjusted to enhance titer...

...biological study without her or her family's knowledge. 3. Fossil Rock Anthem One of Tom's earliest and most...Science with Tom. In this podcast, we learn more about Tom and pick his brain for advice on how to dive... Rap of Henrietta Lacks Students in Oakland rap about Henrietta Lacks - the African American woman whose...Please Don't Kill My Hive Beautifully filmed video about colony collapse disorder.     5. Plasmid Rap ...science standards and communicate science. Learn more about Tom on his youtube channel and at sciencewithtom.com...

...passion and pay attention to the details. Chapter 3 - Answering interesting biological questions using...of our educational resources. Listen to hear all about Professor Goedhart, his lab, and some of the tools...

...degradation of endogenous proteins (notably, BRD2/3/4, CDK9, TRIM24, FLT3, BTK, and ALK) by linking small...with dTAG-13 at doses as high as 20 µM (see Figures 3 and 5 in Nabet et al., Nature Chemical Biology for...degrade any protein. We are particularly excited about the dTAG system as we can use it to evaluate the...

...targeted sequence at the locus of interest. SpCas9’s 3’ NGG PAM occurs frequently in GC-rich genomes, but...100) xCas9(3.7)-ABE 69 (1.4) 43 (2) 21 (3) Not examined 16 (>100) Average percent editing...Blog Learn about Engineered Cas9 PAM Variants Read about CRISPR Base Editing Learn about Homology Directed...editing specificity. We’re excited to learn more about xCas9 - here’s what we know so far! Why do we ...point mutations, also has a strict editing window about 15 nt upstream of the PAM. Even if there’s a PAM...in mammalian cells, finding xCas9 3.7 displayed about the same editing rate as SpCas9 at an NGG PAM. When...are exciting, there’s a lot we still don’t know about xCas9. “I’m not 100% sure xCas9 is going to be flat...

... of the optical fiber into the region of interest 3) Behavioral experimentation I’ll discuss some of the...thoroughly elsewhere (here and here for instance) [2], [3]. Karl Deisseroth of Stanford University , one of ...10.1146/annurev-neuro-061010-113817. PMID: 21692661.  3. Bernstein JG, Boyden ES. Optogenetic tools for analyzing... electrophysiology data too but that’s what collaborators are for). For example, if you study neural pathways...intended brain region and/or cell type (I’ll talk more about this in detail later). But what are the different...of your opsin, but keep you eyes open for posts about the next steps in the coming weeks. Expression of...Addgene. See the Deisseroth lab homepage for details about types of constructs available, instructions for ...

... The collaborative nature of working in industry and how delegating can empower people Chapter 3: How ...therapeutics at Novartis - listen to learn more about Crystal’s exciting career path, to gain insight ...metrics for success. The importance of thinking about the big picture and not just the next experiment...resources in our Mentoring for Scientists Series Learn about Turning Your PhD into a Career with Joanne Kamens...

...Burgess, Shawn., et al. "Questions about NgAgo." Proteins & Cell (2016). 3. "NgAgo: Genome-editing by a DNA-guided...success of CRISPR-Cas9 across multiple model organisms (3) lies in the versatility of its delivery methods. ...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...PMC4343198. Resources on the Addgene Blog Learn More about NgAgo Enhance CRISPR Targeting Specificity with ...

... the lysosome where they are finally degraded (2, 3). Once you've created your fusion protein, it's important...2000. The Cell: A Molecular Approach. 2nd Edition. 3.  CureFFI.org. Cell Biology 04: The Secretory Pathway...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...

...ensure nothing is covering it. Turning on the UV light 3-4 times a week for up to half an hour can help keep...Journal of applied phycology27.6 (2015): 2321-2332. 3. Contamination, Ileana S., John P. S., Basic Methods...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...