...photostability under two-photon illumination than previous GEVIs. Some constructs are also available as viral...kinetics and photochemical characteristics than previous NIR calcium reporters iGECI and NIR-GECO2G, and...reduced sensitivity to pH and calcium compared to previous lactate biosensors, and its affinity is better-matched...temperature sensor that significantly outperforms previous designs. B-gTEMP is excitable under blue light...B-gTEMP itself is a significant improvement over previous temperature indicators, it also demonstrates that...Intracellular Heat Transfer and Thermal Property Revealed by Kilohertz Temperature Imaging with a Genetically...

...suggest that phosphorylation is far more prevalent than previously appreciated. PhosphoSitePlus, one of the... group. Additionally, these mutations are not reversible, and thus cannot reflect the true state of protein...Extensive genomic sequencing of prokaryotic lineages revealed a curious anomaly amongst a subset of methanogenic...mutation of these residues. Due to the limitations previously mentioned, phosphomimetic mutations on the activation...affects protein function, researchers should try to reverse the effects of pSer-incorporation on enzyme function...

...the continued utility of Cre-lox. Check out our previous blog post, Plasmids 101: Cre-lox, if you need ... removes one of the other loxP site variants, preventing further recombination. Brainbow-1.1 scales up...that transient Cre recombination is required to prevent constant inversion of the construct. Sanes and...their protein sequences are highly homologous, preventing design of antibodies specific to each fluorophore...of delicate axonal and dendritic processes not previously visible with Brainbow-1 and -2. The general structure...connectome. Lichtman JW, Livet J, Sanes JR. Nature Reviews Neuroscience. 2008 Jun;9(6):417-22. doi: 10.1038...

...as a nonprofit under the United States Internal Revenue Code 501(c)(3). That means we were formed to benefit... are able to fulfill our mission by generating revenue through distribution of research materials. Our..." is a bit misleading. Nonprofits can generate revenue (as Addgene does through materials distribution... nonprofits should have some level of positive revenue to build reserves so the company is sustainable...The difference is what nonprofits do with their revenue. A nonprofit organization cannot distribute profits...

... news is that this effect is reversible and the pH shift can be prevented. Keep this information in mind...if you’re receiving samples on dry ice - it may prevent you from seeing some unexpected results. Order... 3). However, 30 seconds was not sufficient to prevent any pH change, as we still observed a slight change... allowed the CO2 to leave the tubes, and would prevent the pH change upon thaw (Fig.4). To test this, ...

...entirely too much work for one person, it would take forever, and the end product would likely be poor quality... delegate the giving of feedback or performance review. You can’t delegate tasks assigned to you and you...outcomes, timelines and deadlines. Agree on a way to review project progress. I recommend putting a regular...approachable mistakes will be made that could have been prevented.  Make sure everyone understands that no question... Aren't You Delegating" in the Harvard Business Review. Another excellent blog on delegation tips and ...

...sequencing, revealing that they bind to a much larger subset of protein motifs than previously thought. ...T7 terminators adjacent to the T7 promoters to prevent the incorporation of non-specific nucleotides into...proteomics in model organisms in which it was previously not an option, including Drosophila and C. elegans...Pubmed PMID: 30370338.  SH3 domain collection reveals novel peptide ligand motifs Article contributed...

...your vocabulary! A brief review of base editing tools First, let’s briefly review how CRISPR-derived base...The first base editors revolutionized CRISPR gene editing. Cytosine base editors (CBEs) and adenine base...the pyrimidine bases (C and T). (Check out our previous post on these editors for more details on how ...also called apurinic or apyrimidinic sites and abbreviated as AP sites. To explore this further, the team...bound, keeping the two DNA strands separate and preventing repair based on the complementary strand. This...

...Advanced microscopy, University of Amsterdam. The previous two posts in this series described a practical...between the FPs interfere with native interactions, preventing proper localization in these assays. However,...tendency in living cells. Strikingly, the OSER assay reveals that tagRFP does not act as a true monomer in cells... cells (Constantini, 2012), whereas it was previously concluded to be monomeric based on gel filtration... that are relevant for your research. This will reveal the performance of fluorescent proteins in your...

... titles, authors, year, narrow by article type (review, patent, clinical study, research article etc)....on a new literature search, try to find a recent review article to give you an overview of the topic and...information dense and since it continually builds on previous studies, the reader always enters in the middle...introductions should be fairly focused. Consult a review article for a broader scope of what’s going on ...performed that could influence the results? The peer-review process is not perfect, so even though it is science...

...inputs. This study revealed that these neurons received more inputs than was previously thought. The differences...subsequently higher and higher order neurons doesn’t reveal network connections in a useful way, but rather... tracing is a powerful tool that can be used to reveal brain circuitry, and continue to contribute answers..., Berninger B. Retrograde monosynaptic tracing reveals the temporal evolution of inputs onto new neurons...

...In this study, the labs found that anti-CRISPR prevents Cas9 activity in HEK293 by reducing editing from... Acr samples. Acr proteins can also be used to prevent dCas9 tethering to DNA. Since these early anti-... AcrIIC2 interactions with Cas9 interferes and prevents the correct assembly of the crRNA-Cas complex ...binds to the HNH endonuclease domain of Cas9 and prevents target DNA cleavage (Harrington et al., 2017)....vectors. Having a way to shut off CRISPR-Cas systems prevents the off-target effects and cytotoxicity that result...

...explosion of software tools accompanying the CRISPR revolution. (See omictools for even more.) However, none...step of mutating guide sequences in a way that prevents re-cutting but does not disrupt gene expression...CrispyCrunch is how it mutates guide sequences to prevent re-cutting of the sequence inserted by HDR. Although...mutating a single base pair may not be enough to prevent re-binding. Lastly, the sequence inserted by HDR... target regions Design donor RNA Design primers Review and order reagents Perform your wet lab experiment...

...and I recommend Bustin, 2017 for a more in-depth review.)  Your amplicon should be between 70-200 bp long...ideally located close to either the forward or the reverse primer.  There is a list of useful primer design...efficacy with a standard curve. If relying on previously-generated validation data, be aware that different...transcripts. However, adding too many reagents can prevent your reaction from reaching the plateau stage after...sample compared to your control. But as we said previously, just because it’s used often doesn’t mean it...York 2014Bustin S; Huggett J. qPCR primer design revisited. Biomolecular Detection and Quantification. Volume...

...thus structural epitopes, but can reveal linear epitopes previously hidden within the protein. Created...cross-linking can block the epitope of some targets, preventing your primary antibody from binding (Fig.1). The...cleaner results by not permeabilizing as you’ll prevent binding to any target protein that is still on ...preserve sample morphology, while organic solvents can reveal otherwise hidden epitopes. However, keep in mind...

... unraveling the mysteries of behavior. We’ve previously discussed some of the general considerations ....  What’s involved in locomotor sensitization? Previous research has shown that regions such as the nucleus...transmission leads to locomotion sensitization. (for review see van Huijstee & Mansvelder  2014). Scientists...the locomotion of the animal. This confirms the previously suggested inhibitory mechanism of locomotor sensitization... times are ahead for behavioral neuroscience to reveal insights into the complex relationship between ...

...experimental treatment can be monitored. This can reveal previously unknown functions for genes, and lays the ...of the zebrafish genome was published in 2013, revealing that about 70% of human genes have a counterpart... ways. Zebrafish mutants In the 1990’s, two revolutionary large-scale random mutagenesis screens produced...unlocked novel research into genetic compensation, a previously unknown mechanism of gene regulation (El-Brolosy...

...always to the same extent. As described in our previous Origin of Replication post, DNA replication is...bacteria Before looking at an example plasmid, let's review how E. coli normally replicate their chromosomal...sequence forming this pseudoknot, so its binding prevents the mRNA from folding and blocks RepZ translation...high levels of the lncRNA bind to RepZ mRNA and prevent this pseudoknot from folding, blocking replication...bacterial plasmids. Microbiology and Molecular Biology Reviews: MMBR, 62(2), 434–464. https://doi.org/10.1128/...

...publications. The policy was not an overnight revelation, rather it was the result of consultations between... rationale includes increasing the ability of reviewers (and, once published, anyone) to detect possible...apply different or novel tools and assumptions to reveal findings that the originator was oblivious to, ...called Scientific Data, which aims to act as a peer-reviewed repository for “scientifically valuable” datasets...or table, clicking on the published chart could reveal the underlying data and allow other analyses “on...

...s post and give it a final read Browse list of previous depositors from Boston University Contact Dr. ...that post from Eric before doing anything else. Preventing procrastination Even if you’ve done all the prioritizing...planning in the world, procrastination can easily prevent you from making progress on a task. For instance...to go sit in the tissue culture hood may have prevented me from splitting cells until late in the day ...podcasts. JUST GET STARTED - One Harvard Business Review article on procrastination recommends testing out...