... helped by expanding this team, leading monthly calls, and planning next steps. April Clyburne-Sherin ...first iteration of these workshops took place at Caltech in Jan 2018 and involved presentations and a handout...Not everyone in the group was able to make it physically to instruct the workshop at the conference, but...have the materials and resources to run their own local workshops was a major goal for the team last year...

...widely known of these products is TRIzol® (also called TRI Reagent®, RNAzol®, QIAzol®  and sold by many... within cells or tissues- they don’t actually chemically stabilize the RNA molecules (Allewell and Sarma... MacDonald RJ, Rutter WJ (1979) Isolation of biologically active ribonucleic acid from sources enriched...Guanidinium Thiocyanate–Phenol–Chloroform Extraction. Analytical Biochemistry 162:156–159. https://doi.org/10.1016...

...and can be specifically targeted to a number of different cell types even when systemically administered...with maximum “trunk” space for genetic cargo. Typically, when researchers speak about AAVs, they are actually...ITRs and serotypes The names of AAVs contain two numerical designations, separated by a slash, that indicate...

...Much of today's biological research requires a close examination of specific proteins within a system...without eluting the target protein and needs to be empirically determined. In addition, some metal ion substrates...function. B, The string of histidines in the tag is typically between 6 to 9 residues long. The small size of...Enfors SO, Persson I, Nilsson B and Uhlen M. Large-scale affinity purification of human insulin-like growth...

...protein expression in mammalian cells: easy to use biological tools are abundant! In this post, we will review...that! CRISPR interference (CRISPRi) targets a catalytically dead Cas9 fused to a transcriptional repressor... protein, then these systems need to be site-specifically knocked in. This can be labor intensive and ...and target-specific protein degradation. Nature Chemical Biology. May 2018. 14(5): 431-441. PubMed PMID...

...arXiv.org), but have been only adopted by biological and biomedical scientists (see: bioRxiv.org or medRxiv.org...Preprints can be read freely by anyone and are typically free of charge for authors to post. They can be... be assessed and commented on by readers, typically peers in the field, and updated by the authors over...

...exciting new project aimed at understanding how a critical cellular pathway is regulated. You’ve read all...do you know about the vendor and the vendor’s technical support? An antibody from a vendor that you use...frequently, with a responsive and knowledgeable technical support team, is a safer bet than one from a large... all validation is equal and you should think critically about what data is and is not shown on an antibody...

... Not only is CRISPR having effects on various biological disciplines, the base technology itself is constantly...at non-target sites. Means to decrease these so-called “off-target effects” have included using a Cas9... the Broad Institute, the Joung lab at Harvard Medical School, and the Doudna Lab at Berkeley set out ...afforded by gRNA binding at non-target sites would theoretically decrease off-target cutting. Figure 1...

...plasmids. These are prone to error — both human and biological. Even the most high-fidelity polymerases can ...the correct inserts, tags, promoters, and other critical functional components. Fun fact! Some older plasmids...sequences. These sequences can also be confirmed or theoretical. No matter the level of information that is provided...the raw NGS reads and reassemble using a program called Geneious. Here, the raw reads are mapped against...

... Jesse Zalatan’s lab deposited plasmids for a chemically inducible CRISPRa system in yeast is mediated...Find the yeast CRISPRa plasmids here Reduce the scale of your CRISPR screens by using this lentiviral ...paired guides per construct and allows a reduced scale in screens without any reduction in performance ... channelrhodopsin (C1V1) targeting parvalbumin cortical interneurons is now available as a ready to use...

...choosing antibodies   Complexity is a key factor Biological systems tend to be complex with a variety of ...multiple targets in parallel. In these cases, it is critical that users choose antibodies that are easily distinguishable...staining multiple targets indirectly, you must strategically plan primary/secondary antibody pairing such...Fisher, S. K. (1993). Chapter 15 Postembedding Immunocytochemical Techniques for Light and Electron Microscopy...

...researchers to identify and isolate very specific biological targets. However, as popular and useful antibodies...antibodies, creating a potentially unlimited supply of identical antibodies (See Fig 1.). rAbs are a relatively...has specific binding and has a strong signal. We call the unique pattern of the antibody binding “distinct...TS (2015) The Economics of Reproducibility in Preclinical Research. PLoS Biol 13(6): e1002165. https://...

...things more interesting, we are working in a new technical discipline that nobody understands (synthetic ...thing is opening a pizza joint. Silly ideas (mining local salt deposits to sell at farmers markets), and serious...that survived the literature review, survived technical feasibility analysis, and was a marketable product...and rapidly indicate their presence. They are genetically modified organisms, and GMOs have a bad reputation...

... Liu, 2023). This method uses a prime editor (typically a Cas9 H840A nickase fused to a reverse transcriptase... design of the pegRNA has been shown to play a critical role in the editing efficiency of prime editing...experimentalist, and especially shines once large-scale prime editing experiments come into play. Nicolas...s41587-023-01678-y Li, X., Chen, W., Martin, B. K., Calderon, D., Lee, C., Choi, J., Chardon, F. M., McDiarmid...

...therapeutic strategies to target it, than you typically would at a microbiology meeting. Create an outline...lab mates, and PI can provide feedback on your technical presentation, but those outside of your field ... through the list at the session so you can strategically use your time. Don’t have access to a list of...titles to find the ones you’d like to revisit. Strategically traverse the sea of posters If you come prepared...

...can be long—5 years or more, especially in the biological sciences—whereas industry postdocs have a set...course, you might have less time to explore scientifically since you can’t afford to spend too much time...Unless your academic lab is closely involved with clinical trials or has ties to industry, the drug discovery...industry postdoc, you might find yourself feeling scientifically isolated if no other postdocs are in your department...

...margins of my protocol. While these tips weren’t critical to the experiment, they definitely made my life...qPCR Protocol for more details. Pro Tips When calculating the titer for your AAV, remember to account for...control (NTC) should always be included. While the physical titer generated by qPCR is useful, I test a few...tool for gene expression in mammals Use the PEI calculator for planning AAV packaging transfections Find...

...your gene of interest is difficult to target specifically with a gRNA). Be sure to keep these techniques...sequences are exchanged between two similar or identical molecules of DNA. Gene targeting takes advantage...homology is required, but 6 to 14 kb of homology is typical for targeting constructs. In the example shown ...cells participating in development or other physiopathological events to which the chosen promoter responds...

... 2017. At Addgene, we continually use the Basic Local Alignment Search Tool (BLAST) provided by NCBI. ...BLAST sequence databases from the dropdown menu. Typically, we use the default nucleotide database “Nucleotide...” program for blastn so that you can extract practically every single reliable base pair from your results... the “Align two or more sequences” option to drastically reduce waiting time for search results. Optimizing...

...-length viral genome (Figure 2). While it is technically possible to encode all four viral genes in the... (see Uses of Minigenome System). From a more technical standpoint, a plasmid with all four genes and ... gene by altering the amount transfected (see Technical Tips). Minigenome replication and transcription...compounds which often target the viral polymerase. Technical tips It is important that all of these plasmids...