...with your blocking, first check your antibody spec sheet. Some antibodies work better with specific blockers...include recommended blocking conditions on the spec sheet. If you’re still seeing high background after following...longer incubations at both temperatures if needed. Washes Check your antibody manufacturer for wash recommendations...consider adding a detergent like Tween-20 to your washes — although this can be harsh and inhibit some detection... Inefficient blocking Check antibody spec sheet for buffer recommendations Use a chemical blocking...signaling after each round of stripping* Use a harsher stripping buffer Too little signal Large ...

...yearly planning meetings revolve around two simple worksheets, each of which is filled out separately by the...the trainee and PI before the meeting. These worksheets, summarized briefly below (and available of in...facilitate honest conversation. Goals and Planning Worksheet: Lists accomplishments, research/professional...often give ourselves credit for what we have accomplished. Starting with a recap of accomplishments, from...

...CRISPR-based phenotypic screen datasets. Original published CRISPR screen data is curated and deposited in...in BioGRID ORCS to allow facile queries of published data by biomedical researchers and ready access to... approach serves to maintain consistency with published screen interpretations and allows users to query...curators at [email protected] to submit their published data for rapid curation review and upload. In ...

...wanted to do when I grew up until I was all but finished my undergraduate degree (and I hope I’m not alone...towards a career in science communication, an established field of inquiry with which I was not yet familiar... a scientific background and bring with them established interests and areas of expertise. Perhaps the...strongest aspect of science communication, then, is its sheer versatility in the face of disparate spheres of ...

...2013 there have been 33 CRISPR software tools published and documented by OMICtools. It must be confusing...The first of only three such databases, EENdb (published 1 Jan 2013, no longer available), is a simple ...sgRNA designs. The second database, CrisprGE (published 27 Jun 2015), is noteworthy for its broad scope... content, and ease of access. The third, WGE (published 15 Sep 2015), is more similar to EENdb than CrisprGE...in detail, but that is already covered in the published articles. Instead, I have broken down the tools...

...available. With new fluorescent proteins being published every month, deciding on the best protein for ... to choose fluorescent proteins that can be distinguished from one another and that can be imaged with...common use (particularly not the most recently published ones), they can be a good starting point. In many...abundant proteins or marking compartments. I hope this sheds some light on multicolor imaging with fluorescent...

...Central PMCID: PMC3785459. 2. Krashes MJ, Koda S, Ye C, Rogan SC, Adams AC, Cusher DS, Maratos-Flier E, Roth...induce neuronal silencing in vitro and in vivo (Krashes et al., 2011). The various serotypes available ...by the small molecule clozapine N-oxide (CNO) (Krashes et al., 2011). Viruses expressing pAAV-hSyn-DIO-hM4D...

...scientific communication needs. Going to present some unpublished results at a conference? Post your working manuscript...scientific community. Most preprints are eventually published in journals after formal peer review, with the... as the source or were deposited when the now-published article was a preprint. Depositing plasmids that...bioRxiv. https://doi.org/10.1101/833400  Stern BM, O’Shea EK (2019) A proposal for the future of scientific...

... easily applied to multiple dishes, remember that handling multiple dishes will take longer and may increase...scaling up by either increasing the number of 10 cm2 dishes or increasing the vessel size. Opting for a larger... opt for simply increasing the number of 10 cm2 dishes. While this option is attractive because the same...

...affinity: stronger binding antibodies will require harsher stripping to remove, so start with the weaker binding... membrane will remove some of your sample —and harsher stripping means more sample loss. If you are still...a recipe like the ones shared by Bio-Techne, ThermoFisher, or Millipore Sigma. We’ll share the basic composition...solutions below. Alternatively, companies including ThermoFisher, Millipore Sigma, and Azure Biosystems offer...

...performance in our system and yet none of that data was published.    Fig. 1: Testing of three different ... for your system or to successfully replicate published works. Of particular importance to these efforts...build on others’ results is available not through published manuscripts, but through word of mouth or old ...asked your team members or colleagues: How much unpublished AAV data do you have sitting in computers and...

...This post was originally published on the Quilt Genomics Blog and is republished here with permission. Quilt...enhancers, check out the Appendix. Conclusion: Shedding light on dark matter Precise knowledge of which...identified 23-mer gRNA. Off-target determination was established with Bowtie2 using the parameters first described...

...factors OCT3/4, SOX2, KLF4, and c-MYC is the established model for reprogramming somatic cells. A common...Polycistronic cassettes that contained KLF4s had overall diminished KLF4 expression levels; this altered stoichiometry...as inhibitory GABA-ergic interneurons, Gordon Fishell’s lab has developed a new AAV approach using regulatory...to Choosing the B(right)est Fluorescent Protein. Shen Y, et al. PLoS ONE. 2017. PubMed PMID: 28241009 ...independent of another TTAGGG binding complex, shelterin. Swapping the DNA binding domain of another telomeric...

...centrifugation or a magnet. 4) Unbound proteins are washed away. 5) The target protein is eluted.     ...and as in the first method, unbound proteins are washed away and the target protein eluted.  Immobilization...providing a solid support while unbound proteins are washed away.  While both Protein A and Protein G have ...because they do not require centrifugation between washes which can cause loss of beads, break weak protein...

... to only do the dishes once a month, resulting in a dangerous mountain of dirty dishes or you could choose...choose to do the dishes after every meal and be reassured that your dirty dishes won’t tumble out of the...

...chamber is divided into two halves that are distinguished visually through different patterns and the ...recently studied in detail thanks to optogenetics. In Goshen et al, inhibition of the CA1 region using HR was...your own neuroscience sub-discipline that has published using the technique and ask if they would be willing... opsins." Nature methods 9.2 (2012): 159-172.5. Goshen I, et al. Dynamics of retrieval strategies for ...

...publications and funding. If any of these three pillars crashes, the lab's success and the careers of its researchers...not enough. A 2013 study showed that out of 238 published scientific articles, only 46% could be reproduced...adopting simple note-taking tools, others using excel sheets or organizing folders in complex ways, there are...everything in one place. From pictures to excel sheets, researchers today generate vast amounts of digital...

... Additional resources ThermoFisher Scientific: Biotinylation ThermoFisher Scientific: Biotinylation...more information on ABC and IHC check out Thermo Fisher Scientific's guide. There are an array of other...which biotinylation reagent to use check out Thermo Fisher Scientific's Biotinylation Reagent Selection Tool...

...cytoskeleton and cell membrane dynamics. First, a quick refresher.  The tools below use the following optogenetic...reversible. Late last year, the Akhmanova Lab published a new optogenetic tool for localized microtubule...the membrane itself? The Baskin Lab previously published OptoPLD, an optogenetically controlled phospholipase...available for controlling cell dynamics. Need a refresher on VVDfast, iLID, LOV, CRY2-CIB, and other optogenetic...

...sequencing or other readout. Cellular memory is accomplished by the use of sequence-specific enzymes (e.g...also available, N1-LOC8R.  These plasmids are unpublished Unnatural amino acid incorporation in mammalian...detection of neuronal action potentials. The Cohen lab wished to improve on the Arch indicator and their collaborators... that their sgRNA-RNA fusions were functional, Shechner et al used the constructs in their study to target...contiguous with the sgRNA core (see image above). Shechner, et al. Nat Methods. 2015. PubMed PMID: 26030444...regulation in live cells since their introduction by Sherf and Woods in the nineties. However, reliance on ...guess what? pXPG is now available at Addgene. Sherf, et al. 1994. Promega Notes Mag. Giannakis, et...