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...passed down through generations (Figure 1). These so called “plasmid addiction systems” require dividing cells..., plasmid addiction systems have been used as biological tools for a wide range of applications in molecular...become a powerful tool to address a diverse set of biological inquiries. References Ogura, Teru, and Sota...vaccine strain CVD 103-HgR." FEMS Immunology & Medical Microbiology 57.1 (2009): 69-79. PubMed PMID: 19678844...

... defective gene, Xm, the sequence of which is identical to the correct gene, Xw, except for a single base...precisely changes a single base with an efficiency typically ranging from 25-75%, while the success of precise... in the DNA by cutting only a single strand, dramatically reducing the likelihood of detrimental indel...known 'DNA adenosine deaminases' which would theoretically give rise to an adenine base editor. A massive...

...possess cell walls. As a result, Mycoplasma are intrinsically resistant to this class of antibiotic. Acquired...When antibiotic resistance testing is requested clinically, this involves growing a standardized amount ...certain antibiotic were chosen for treatment. This is called a Kirby-Bauer disk diffusion test or Kirby-Bauer...-trusted method that is frequently used in the clinical laboratory. What can we do to stop the emergence...

...contamination. For those labs that prefer a more economical mode of detection, Uphoff et. al. outline a do...engineered with non-native envelopes, a process called pseudotyping. Lentiviruses pseudotyped with the...virus. To circumvent this problem, transductions typically employ the positively charged polycation, polybrene...lentiviral and retroviral vectors.” Human Gene Therapy Clinical Development 25.3 (2014). 175-85. Pubmed PMID: ...

...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 ...

...regulators. But the speed is impressive on a larger scale, too. The first paper reporting genome editing by..., 2016). Advancing from this invention to KJ’s clinical application in under a decade is an astounding... safety of KJ’s experimental treatment, it was critical to reduce off-target editing as much as possible...P. (2025). Custom CRISPR-Cas9 PAM variants via scalable engineering and machine learning. Nature, 643(...

...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...

...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...

...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...

...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...

...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...

...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...