...secondary antibodies instead (B).   Since a polyclonal primary antibody contains a mix of different...antibodies used for detection bind to non-target immunoglobulins naturally present in the tissue sample. There...antibody fragments to block the endogenous immunoglobulins and reduce the background staining but they...Biological systems tend to be complex with a variety of proteins interacting. Experiments tend to be equally complex...

...mutation in the evolved TadA domain. Because ABE7.10 contains a heterodimeric fusion of wild-type TadA and TadA7.10... the evolved TadA region. The ABE8.x-m version contains only the evolved TadA and is ~500 base-pair smaller...evaluate further.  The creation of accidental insertions and deletions (indel) by the catalytically impaired...more than 90% compared to ABE7.10. Choosing Cas9 proteins to increase the targeting scope of adenine base... Cas9 in the ABE complex with PAM-variant Cas9 proteins: the S. pyogenes NG-Cas9 (PAM: NG) to create NG-ABE8...

...challenge is finding relevant papers. The latest installment of the Early Career Researcher Toolbox will highlight...search that’s constantly updated. This Twitter list contains >300 Twitter literature bots, including a bot ...AI-backed literature search developed by the Allen Institute for Artificial Intelligence. Semantic Scholar ...Initiative. Meta is not a traditional search engine. Instead, you create your own research feed by entering ... papers are published, similar to Facebook or Instagram. Key features Setting up a personalized feed is...

...first make your poster. While a poster generally contains the same sections as a primary research article...in your poster. This is a section where you can insert your references and contact information, and it...conclusions, and acknowledgements sections are inserted. The advantage of this style is that it will display... a poster vertically rather than horizontally. Instead of displaying your information as columns, you ... poster, written and graphical information are inserted into larger circles or ovals, that are arranged...

...distinct proteins and result in unclear transcriptional consequences (Sprujit 2013). Enzymatic domains tethered...are a number of drawbacks to this approach: TET proteins are not demethylases; they are dioxygenases that..., Charlton et al. 2020). Most importantly, TET proteins affect transcription independent of their enzymatic...overlap the CpG of interest. Having the CpG directly inside the gRNA sequence is a great guarantee that DNMT1...

...use a standard strain, like DH5α, grown at 30 °C instead of 37 °C. If you’re unsure about the integrity ...prefer to store freshly harvested virus at 4 °C instead of storing it briefly at -80 °C. If troubleshooting...option may be to avoid freezing altogether, and instead to use the virus immediately upon harvesting. Interestingly...appear as a white pellet. The pellet can then be rinsed and resuspended in cold sterile PBS.  Alternatively...of ensuring viral production is to perform PCR against components of the viral vector using the harvested...

...Director of the National Human Genome Research Institute (NHGRI) of the NIH, started off with the first...will lead to advances in human health. It was inspiring to hear how the genome project has already had...enzymatic machinery (i.e. RNA replication with no proteins). The chemistry is complex and this work requires...desribed how P. furiosus has systems to defend against both DNA and RNA viruses - 2 cas systems regulate...system. His lab is also exploring the use of cas9 proteins from different species for genome engineering....

...—an optical insulator called a ferrule (below). The fiber optic cable/ferrule is inserted into the target...stimulation guarantees that the majority of the opsins will receive sufficient quantities of photons to...although you may be able to purchase one pre-made instead). Laser stimulation         In the video above...: closed/light receptive, open, closed/light insensitive. If you deliver a continuous stream of photons...

...variance of size of plasmids in your library (Are the inserts of different sizes?), and the desired coverage ...100x coverage of your library, and your library contains 5,000 plasmids, your goal for your transformation...replicate faster. For this reason, if your library has inserts of different sizes (such as a transcription factor...library is of a smaller size with a single size insert, liquid media can be faster and easier to work ...protocol paper by the Zhang lab, and the Broad Institute’s general pDNA Library Amplification protocol are...

...practice: Rehearse your presentation with people inside and outside your field. Your colleagues, lab mates... tack up your poster seconds before the session begins. Plus, you’ll have extra time to practice at the... in to the next figure. Remember your outline: Instead of getting bogged down in the minutia or coming... practicing with your colleagues, ask them to brainstorm potential questions you may receive. People often...in to thirty-minute-long presentations, which constrains the number of posters you can see. If you are...

...pastoris) because it secretes fewer of its own proteins, making the purification process simpler. Due ... the production of Fab fragments where variable chains are secreted into the periplasmic space to be oxidized...SDS-PAGE gel. Once stained, the heavy and light chains become visible alongside your ladder and a protein... density of the antibody bands can be measured against the protein standard and the concentration may ...Any bands on the gel beside the heavy and light chains can also be measured. Contaminating bands lower...

...sensors. Given the expanding palette of fluorescent proteins and the availability of powerful imaging technologies...modules used in this method include fluorescent proteins, promoters, enhancers, polyadenylation signals...combined to create a single polycistronic 15 kb insert. The final construct was then introduced to HeLa...later combine all four constructs into one 15 kb insert. Table 1: MXS Constructs for Labeling Cellular ... fluorescent proteins...

... for instance, a recent preprint released by the Boehm and Beroukhim labs at the Broad Institute of MIT...MIT and Harvard, Dana-Farber Cancer Institute, and Koch Institute, where an exploration into the effects...introduced an ancestry bias into their experiments instead of a true correlation.   It turned out there was...

...previous books focused primarily on fluorescent proteins and their applications, tackles much broader issues...from mildly surprising to genuinely shocking. Mainstream media outlets have addressed CRISPR frequently...science produced by the quieter, more reserved Franklins and Prashers will find it increasingly difficult...Addgenies Joanne Kamens, Marcy Patrick, Aliyah Weinstein, Fabienne Furt, and Quintin Marcelino for their... Eric J. Perkins...

...DNA N-glycosylase (UNG) enzymes. Various other proteins included in different CGBE tools allow the researcher...a similar tool shortly thereafter, using mouse instead of human AAG (remember, MPG and AAG are the same...a more specific A → C base editor (ACBE), they inserted the deaminase into the C-terminal domain of Cas9...excises inosine can be modified to target guanosine instead, perhaps analogous engineering of UNG, which excises...its homologs) has more than one name. Indel Insertion/deletion A type of unwanted DNA edit. AYBE, ...

... of all the proteins in the class that showed the relative sizes and the conserved domains. This information...highlighted a few dozen things that I didn’t know about. Instead of reading a paper and getting my bearings, I would...summarized how frequently each member of a class of proteins was mutated, as reported in various studies. We...sense of the similarities and differences among proteins in the class. Offer your perspective It doesn...

... thanks to our guest blogger, Jessica Roginsky. Jessica Roginsky leads the scientific support team at ...This article was contributed by Jessica Roginsky, Scientific Support Lead at Synthego. Article source...genomes that have been successfully modified with insertions or deletions (indels) and then characterizes ...each bar of the indel plot shows the size of the insertion or deletion along with the percentage of genomes...

...collaborations. During my graduate studies at Karolinska Institutet I started working with other groups. First...minutes. Get to know your collaborators from other institutions personally on conference calls and make sure...learning and, in my experience, mistakes and insecurities are more easily excused. When you reach out ...Collaborate: An Imperative for Graduate Students" from Inside Higher Ed Additional Resources on the Addgene Blog...

...cassette contains the major structural gene gvpA marked with EBFP2 and the second cassette contains the P2A-linked... be easily expressed heterologously by bacteria inside the body. So we screened gas vesicles gene clusters...bARGSer-expressing E. coli. The GV-specific ultrasound signal inside the tumors was overlaid with the anatomical B-mode...which seemed overexpressed compared to accessory proteins. Excess gvpA plasmid added into the transfection...

... This post was contributed by Ryan Watkins, Professor at George Washington University and developer of...in scientific journals flies under the radar and gains little notice. A 2009 research study found that ...influenced by research in other disciplines. But there remains a great deal to learn from the science of others...platforms (such as social networking, visual appeal, instant sharing across multiple other sites), I created...and let the world know about your science! Ryan Watkins is a professor at George Washington University ...