We narrowed to 1,041 results for: ins
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TypeBlog Post...for every assay. Antibodies that are generated against a linear antigen tend to work very well for applications...application. Take immunohistochemistry (IHC), for instance. While an antibody may be validated for IHC, not...region highly conserved across species? If the proteins are highly homologous between your target species... each isotype or species to easily discern the proteins in your assay. Narrowing the field After working...compare the staining patterns of multiple antibodies against the same target; tagged protein strategies that...
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Announcing the Winners of the 2021 Michael Davidson and Roger Tsien Commemorative Conference Awards
TypeBlog Post...awards to early career scientists using fluorescent proteins in their research. These awards help early career...biosensor fuses MPST to roGFP2. Because roGFP2 contains two cysteines near the chromophore that are susceptible... to better understand the processes that happen inside the cell,” says Pedre Pérez. Figure 2: ...changes following opioid withdrawal, providing insight into new methods for treating patients in recovery...indicator which had improved performance in live mouse brains. Currently, she is using the improved voltage indicator... -
Plasmids 101: Golden Gate Cloning
TypeBlog Post...destination vector + insert may spontaneously religate, this transient construct retains functional Type IIS...in the final construct. The destination vector contains sites with complementary overhangs that direct...complete with helpful tips and tricks, see The Sainsbury Lab website or Engler & Marillonet. Golden Gate...technology has adapted Golden Gate cloning for inserting the appropriate oligonucleotides specifying a ... -
Educational Planarian Tools for Teaching Developmental Biology Techniques
TypeBlog Post... they bind to a class of Argonaute proteins. The Argonaute proteins remove one strand of the siRNA and...cDNA), and then use PCR and molecular cloning to insert the cDNA into the desired plasmid backbone. The...the process is a little more complicated. These strains are more picky eaters and refuse to eat liver paste...Sánchez lab protocol also provides step-by-step instructions for the “the bravest and more curious” students... -
A Guide to Designing a Scientific Poster: Content, Layout, and More
TypeBlog Post...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... -
A New Generation of Adenine Base Editors Improves Editing in Primary Human Cells
TypeBlog Post...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... -
Simple CRISPR-based Epigenetic Editing: dCas9-directed DNA Demethylation
TypeBlog Post...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... -
Antibodies 101: Choosing the Right Isotype
TypeBlog Post...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... -
Early Career Researcher Toolbox: Finding Relevant Papers
TypeBlog Post...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... -
5 Tips for Troubleshooting Viral Transductions
TypeBlog Post...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... -
Reaching out to China: Canton Nucleic Acids Forum (CNAF) 2015
TypeBlog Post...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.... -
The Materials Science of Optogenetics Experiments
TypeBlog Post...—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... -
The Scientific Conference Poster Session: Tips for Success
TypeBlog Post...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... -
Pooled Library Amplifications
TypeBlog Post...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... -
Capturing a Moment with Marc Zimmer’s “The State of Science”
TypeBlog Post...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... -
MXS Chaining
TypeBlog Post...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... -
Antibodies 101: Producing Recombinant Antibodies
TypeBlog Post...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... -
Uncovering Ancestral Bias in CRISPR Research
TypeBlog Post... 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... -
Analyzing CRISPR Editing Results with ICE from Synthego
TypeBlog Post... 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... -
Transferable Skills Guide: External Collaborations
TypeBlog Post...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...