...Professional Career Coaching can be an excellent tactic for scientists making a career shift or who wish...of this to get the information you need to choose well. What are their qualifications? Do they have certifications...does not endorse any of these coaches and I can't tell you which one will be a match for your needs, but...Consulting is part of that group and she is especially excellent in coaching for communication skills. Industry...Industry transition is also an area of specialty for Michelle Capes at Adeptify Me. While based in Madison, ...

...currently doing this using two approaches: single-cell RNA-sequencing to identify neuronal class and deep...Drosophila and zebrafish articles published there as well.   Worm Development Dynamics Database (WDDD) The...WDDD aims to acquire quantitative data on all the cell divisions of the C. elegans embryo when all essential...Wang et al., 2018).” With the split system, only cells that have both halves of cGAL can drive expression... Alyssa Cecchetelli...

... question about what should go into a cell, hover over the cell and instructions will pop up. When you...spreadsheet (Figure 2) and are described in Table 1. Some cells have drop-down options for you to choose from, while... into the appropriate full and partial sequence cells. *Note: If you believe you need to deposit a collection...Relevant Mutations Indicate the amino acid change as well as any functional consequences of that change. For...

...plethora of antibody options for your experiment (all well validated for your application, of course!) When...the isotype, and subclass affect my experiment?” Well, it depends on the application and target. If you... blot and probing a single target that expresses well, then probably not. In this case, take a look at...degree of affinity, then species choice could very well matter. Take rabbit versus mouse monoclonal antibodies...for Light and Electron Microscopy. In Methods in Cell Biology (pp. 283–310). Elsevier. https://doi.org...

...encode proteins. They play important roles in many cellular and regulatory processes, but much of their functionality...RNA (sgRNA) to create a double-strand break works well because very small deletions and insertions in the... should be able to devise an experiment that can tell you more about its function.  Resources and references...screening for functional long noncoding RNAs in human cells by Cas9 targeting of splice sites. Nat Biotechnol...non-coding RNAs with the CRISPR/Cas9 system in human cell lines. Nucleic Acids Research. 2014;43(3):e17. doi...

...better than a nitrocellulose membrane. If you have already run your blot using nitrocellulose, you may decide...protein of interest in a complicated solution like cell lysate. But they can be a lot of work — and what...instructions for the membrane you are using. Dry nitrocellulose membranes are brittle, so proceed with caution...to solutions. It is also volatile and toxic (and smells bad!), so be sure to use it under a fume hood. ...

...since the lab was highly focused on protecting intellectual property, it worked away in isolation. It published...joint. Silly ideas (mining local salt deposits to sell at farmers markets), and serious ideas (how to stop...color-changing flowers was a great business idea as well, and they thought we could be taught. They invested...Biochemistry Department at the University of Manchester as well as the Biology Department at CSU. When not pursuing...enjoys spending time with her husband and two Jack Russell Terriers. More SynBio Reading Read more Addgene...

... in mammalian cells Hu et al. compared xCas9 activity with wt SpCas9 in mammalian cells, finding xCas9...ABE7.10, as well as improvements at NG and GAT PAMs. xCas9 3.7 Activity in Mammalian Cells Construct... percent editing efficiencies (edit frequency in cell population measured with NGS) are shown as percentages...

...doing a brief academic postdoc. Why did I switch? Well, my long-term goal was to enter industry since I...” that you’ve sold out. You’ve given up your intellectual freedom and integrity. (Not the case, as I’ve...s ability to manage others and work as a team as well as their scientific abilities. At AstraZeneca, scientists...who get postdocs and do not support or manage them well are simply no longer allowed to have postdocs. In...Scientist at AstraZeneca. She has a background in cell and developmental biology and is applying this towards...

...overexpress YFG in a Drosophila cell line. Knock downs in Drosophila cell lines are extremely effective....days after setting up the genetic cross). You can tell when pupae are ready to hatch the next day because...person. Animals are diploid except for their germ cells (sets of genomic material are contributed by mother...various fly reagents on the Bloomington website as well. Additionally, they have useful information on topics...

... for Mapping Multiprotein Complexes in Human Cells.” Cell Rep. 13: 621-633. PMID: 26456817. Find plasmids...Chromosomes). Using their PITCh plasmids, GFP knock-in cell lines can be created in about a month and a half...locus-specific gRNAs Step 4: Select for puromycin resistant cells Step 5: PCR amplify and sequence the locus to verify...crispr.mit.edu. They tested their system in HEK293 cells, integrating the GFP-Puro cassette into the FBL ...

...encoded tools, such as microbial opsins, to control cellular activities using light. In 2015, scientists combined...activator domain (VP64 or p65). Upon expression in the cell, the dCas9-CIB1 fusion binds to the target DNA sequence...activator domain VP64. Using this system in HEK293T cells to induce expression of human IL1RN, the researchers...protein with three red fluorescent tags in U2OS cells. Upon light activation, dCas9 is released within... silencing of transferrin receptor CD71 in HeLa cells.   Comparison of photoactivatable CRISPR strategies...Spatiotemporal Control of CRISPR/Cas9 Function in Cells and Zebrafish using Light‐Activated Guide RNA. Angew...

...machinery, and could thus be used in non-dividing cells. Cas13 enzymes also don’t require a PAM sequence... RNA knockdown. In initial tests, Cas13b from Prevotella sep. P5-125 (PspCas13b) yielded 62.9% average... a maximum of 28% editing efficiency in HEK293T cells, as assessed using RNA-seq. Since the REPAIRv1 machinery...including reversibility and use in non-dividing cells. With the expanded targeting ability using both ...Functionally Orthogonal Type VI-A CRISPR-Cas Enzymes." Mol Cell. 66(3) (2017):373-83. PubMed PMID: 28475872. Find...

...Nanobodies are like tiny antibodies which work just as well, if not better, than antibodies for all of the above...structure and function compare to IgG antibodies, as well as how to produce them for use in your lab. Structural...triple colocalization is possible even when staining cells with three IgG1 antibodies. Antibodies were individually...workflow yields similar localization patterns as cells stained for one target (compare nanobodies staining...–rabbit IgG secondary nanobodies. The Journal of Cell Biology. PubMed PMID: 29263082 Find plasmids from...

...not generally in a cell’s best interest to overexpress a protein. Energy and cellular resources are being...being spent to make something the cell doesn’t need to make. Eukaryotes and some bacteria deploy proteosomes...proteosomes to degrade what the cell might consider junk protein. Though there are a number of chemical and...same manner as it would have in the context of a cell. FLAG tags can be cleaved by enterokinase, which...

...day care and school if you tell them they will enjoy it. If you cry and tell them they will miss you, of...having anyway) for enjoying your career and life as well as your family and friends. If there is any guilt... way more fun. And despite those who may wish to tell you so – towel corners do not need to match up exactly...example, say no to some of those birthday parties. Tell your mom you can’t make it to dinner this week. ...

...particles that can transduce cells. Infectious titers are typically quantified by cell transduction assays. Wild-type... on three important factors. What can AAV titers tell you? Physical titer: the concentration of viral...arrowhead). Scale bar = 100nm. Image courtesy of David Bell and Svetla Stoilova-McPhie, Center for Nanoscale...Briggs HA, Gallichotte EN, Ruf IK, Hindson BJ, Vessella RL, Tewari M. Absolute quantification by droplet...

...Specifically, roles in technical support make excellent use of the skills developed by life science PhDs...issue was with a custom-made reagent to manipulate cells (manufactured by one of the larger biotech companies...patient enough to find the real issue, you will do well in a technical support role. What general characteristics...go wrong in a lengthy protocol. And customers may tell all sorts of details about the application, just...about novel developments in the fields of gene and cellular therapies.     Additional Resources on the Addgene...

...how do proteins X and Y regulate the Z pathway?" A well-defined outcome makes it easier for the search committee...example, "while it is possible to grow liver stem cells and liver-like mini-organs or organoids, neither...combine bioengineered scaffolds with patient stem cells to generate compatible functional livers.” Why are...Health Sciences University, did his postdoctoral fellowship with Jennifer Lippincott-Schwartz at the National... optimization of fluorescent proteins,  and live cell imaging approaches. Erik is also an avid long distance...

...off-target effects. The flaws of RNAi screens are well characterized and strategies exist to control for...delivered as a pooled library via a lentiviral vector to cell lines that either stably expressed Cas9 or that ...Hart et al, Bassik et al). The library performed well in both screens, with the growth screen having a...almost all genes involved in the production of a cell surface glycan that’s required for ricin uptake....reveals pathways underlying ricin susceptibility." Cell 152.4 (2013): 909-922. Pubmed PMID: 23394947. Pubmed...