...on DNA, or are enzymatically active. For this reason, recombinant proteins are typically expressed in E...medium. Induction timing is important, as you typically want to make sure your cells have first reached...your expression conditions by setting up a small-scale expression experiment to test variables such as ...

...ICC or a tissue in the case of IHC. IF is used specifically when we visualize the antibodies using fluorescence...counterstaining, mounting sample on slides, and confocal   Figure 2: High-level overview of an ...controls to show that your antibodies are binding specifically. For positive controls, aim to use samples that...in an organism that is underrepresented in the biomedical research community. You may have to take some...

...control over where mutations are introduced, and scalability (see Supplementary Table 1 of Wrenbeck et al)...stretches of sequence could be used. However, a practical limitation is sequencing depth and cost. You need...Measuring the Activity of Protein Variants on a Large Scale Using Deep Mutational Scanning.” Nature Protocols...scanning: assessing protein function on a massive scale." Trends in Biotechnology 29.9 (2011): 435-42. PubMed...

... regions of homology at their ends, which are typically created by PCR. Then, the fragments are incubated...competent cells. The success rate of this reaction is typically fairly high, so there is usually no need to screen...the bacterial chromosome (up to 100 kb) were specifically cut out via CRISPR and then assembled into a...amplify the fragments of the chromosome, which was technically challenging. Other homology based technologies...

...desired location of edit; efficiency decreases dramatically when the cut site is >30nt from the proximal... constraints are even more exquisite for the so-called base editor Cas9, which makes DNA changes in the...nuclease-mediated off-target effects.” Further, large-scale datasets of hundreds of genetic screens using genome-wide... sequences targeting a gene to score in order to call that gene a hit.  This is an exciting time for functional...

...simple in C. elegans and bacteria. But CRISPRing typical mouse models, including creating Cre-dependent ...on both sides and the T7 promoter on one end. Typically, Easi-CRISPR works best with insertions of 2 kb...models varies from 8.5-100% of pups born, with typical efficiencies of 30-60%. Quadros et al. attribute...2.5x more animals than i-GONAD and is a more technically difficult process. Overall, the main drawback...

...facilitates the integration of the transposon specifically at ‘TTAA’ sites randomly dispersed in the genome...re-integration from the piggyBac™ donor, which is typically found with the wild-type piggyBac™ transposase..., including making transgenic cell lines and genetically-modified cell lines. Furthermore, the use of ...cell engineering team.     Mathew Goodwin is a Technical Sales Specialist at Transposagen. As a gene editing...

...the details. Chapter 3 - Answering interesting biological questions using microscopy. Chpater 4 - Research...Amsterdam). He develops, characterizes and uses genetically encoded fluorescent probes. You can follow him...Plasmids for FRET Find Plasmids for Subcellular Localization ...

... BioRender.com.   Cas9 has two major lobes, called NUC (for nuclease) and REC (for recognition). The...recognizing the PAM on DNA, the CTD is also sometimes called the PAM-interacting (PI) domain. The guide RNA ...conformational state of the HNH domain also allosterically controls RuvC activity through a hinge region...Visualization, and Informatics at the University of California, San Francisco, with support from National Institutes...

...African American woman whose cervical cancer cells became a mainstain of biological study without her or her...fossil rocks and their importance in understanding biological history. 4. Please Don't Kill My Hive Beautifully...

...important immune cell type, in more complex and physiological microenvironments. These screens have primarily...molecular identifiers, which can improve the statistical power of in vivo screens. Figure 1: Using...Fellow in Arlene Sharpe’s laboratory at Harvard Medical School and is interested in improving CD8+ T cell...

...alternatives for some of the harmful chemicals in your lab. These chemicals are not only better for the environment... resources about this, and even has a list of biological samples that are stored long term at -70 °C. ...

...plasmids available at Addgene. We will still periodically focus on specific CRISPR plasmid tools more ... Feng Zhang’s lab recently developed a platform called SHERLOCK (specific high-sensitivity enzymatic reporter...will help researchers map transcription factor localization and binding via ChiP-seq. Learn more about...

...papers detailing Nobel Prize-winning research typically weren’t among the most cited. And, as scientific...online, surely times have changed. Altmetrics.org calls to expand our view of what impact looks like and... aren’t enough. Scientists are sharing what they call “raw science” – data sets, code, and, I think it...

...protein present on the virion surface – making it critical for viral attachment and fusion. Once inside a...glycoprotein’s role in infection also makes it a critical component of vaccines and the target of neutralizing...established Ebola Genome Portal at the University of California, Santa Cruz, offers sequence data on five strains...

...multiple timepoints. When the target of interest is critical to cell survival or proliferation, an additional...a flow cytometry experiment. But validation is critical to ensuring accurate interpretation of your results...to start with a screening procedure in easily genetically modified cell types (top). The specific approach..., A. M., McPherson, P., & Laflamme, C. (2023). Scaling of an antibody validation procedure enables quantification...

...few years ago, researchers introduced what’s been called a major breakthrough in cloning technology with...cassettes onto a single T-DNA in subsequent rounds, dramatically reducing the time required to generate multi-construct...multi-construct transgenic plants. In the most practical terms, what GreenGate can do is “save you half a...

...promoter. It can be used to drive Cre expression specifically in neurons. AAV Delivered Cre - A useful tool...Indeed, this AAV provides you with a means to specifically activate expression of any Cre-dependent gene...Additionally, you could use the retrograde form to specifically express Cre in neurons with axons that extend...

...sequence in live cells. This technique uses catalytically inactive CRISPR endonuclease (dCas9) fused to...scientists. You can find all of our August posts in chronological order here: Site Directed Mutagenesis by PCR...Single Base Editing with CRISPR Cas9 Activators: A Practical Guide 10 Steps to a Perfect Science Talk Developing...

...meet interesting people doing interesting things. I call this building relationships because networking has...or a particular position at their company, do NOT call this an Informational Interview. If you do this,... if it is not possible consider a phone or video call. If you do decide to use Skype, make sure to follow...