...on DNA purification without a kit that outlined several reasons why doing something without a kit has advantages...everything, including your skin. Here are some general tips for working with RNA, even if you are using...method, so biotechnology companies have marketed several products that help to further streamline this process...quantitative PCR to assess gene expression. There are several things that researchers can do to combat this issue...and proteins. Lastly, using a DNase (there are several DNase enzyme products on the market to choose from...

... cloning, you’re probably aware that there are several methodologies currently available for approaching... and Gateway® cloning technologies, as well as several others. Each method is unique and relies on specific...to propagate because CcdA wasn’t available to counteract the toxin. If, however, an investigator were ...answer lies in special strains of E. coli that tolerate the expression of the toxin gene. One such strain...plasmid CcdB protein involves poisoning of DNA-topoisomerase II complexes.” J Mol Biol. 1992 Aug 5;226(3)...

...studied a synaptic protein that is linked to neurodegeneration. The level of this protein is decreased in...method for the visualization of protein-protein interactions in situ. In the standard PLA assay, primary ...endogenous proteins of interests, suspected to interact, are added to a cellular sample. Both antibodies...visualization of the location of the protein-protein interaction. To study protein levels in situ, the Schumann...protein plasmids Use FRET to study protein-protein interactions Deposit plasmids with Addgene! ...

...genome editing in large-scale has led to the rapid generation of genome-wide gRNA libraries that knock out ...themselves incredibly useful for the efficient generation of individual (and multiple) genetic perturbations... al., 2015; Wong et al., 2016). CombiGEM uses iterative one-pot reactions with straightforward restriction...functionally characterize combinatorial gene knockouts generated via multiplexed gRNA expression (Wong et al., ...gRNA targeting sequences, you can then readily generate your barcoded gRNA library sequences via oligo...

...recognize multiple isotypes from a species, like a general anti-mouse antibody. Others recognize specific ...better choice for a mouse monoclonal IgG2a than a general anti-mouse IgG. In a broadly specific secondary...different species.      Table 1: Isotype considerations when choosing antibodies   Complexity is...tend to be complex with a variety of proteins interacting. Experiments tend to be equally complex with ...narrowly specific secondary antibodies rather than generally reactive ones. And make sure to check for any ...

...interviews, however, were more structured. I met with several individual and/or small groups, usually starting...interviewer,” you can find lists that have lots of general questions about job responsibilities, performance... an opportunity to demonstrate technical and transferable skills that make you qualified for the position...position. Highlight the outcomes or deliverables of your work, such as improving a process or procedure, ... pursuing a particular line of research is a deliverable. Find 10 steps to a perfect science talk Be prepared...

...Collection at Coriell. More information and images generated from these cell lines can be found in the Cell...functional aspects of the protein gleaned from the literature and engagement with researchers who study the...find out more about gene edited hiPS cell lines generated using the Allen Institute for Cell Science Plasmid...Drosophila cells, and C. elegans. In addition to generating fluorescent protein fusions, plasmids are available...find out more about gene edited hiPS cell lines generated using these plasmids, please visit the Cell Catalog...

...dimeric nature can interfere with some applications. Several labs have been working to produce monomeric versions... under CC-BY license.   The Miyawaki lab generated a series of StayGold mutants and identified a ...mentioning that Kiryl Piatkevich and colleagues generated yet another monomeric StayGold variant (Piatkevich...primates. The authors used a multispecies approach to generate and screen a library of AAV9 variants (Chuapoco... brains. Figure 3: Overview of library generation and screening that led to identification and ...

... preparation of the tissue sample, several washes, and temperature changes to visualize the expression...to study gene expression and functions of key regeneration-related genes in four inexpensive and commercially...linearized DNA templates which they can easily generate off of a circular plasmid via PCR. Typically, ...expression is activated through the  induction of T7 polymerase encoded in the E. coli genome (usually controlled...

...Alberts et al, PNAS 2014; Taylor, eLife, 2016). Several Future of Research (FoR) symposia held across the...and Molecular Biology/ FoR Hack Day during which several projects were initiated. Recent developments promise...the passing and upcoming implementation of the Federal Labor Standards Act (FLSA) which mandates that ...different departments across the university and alterations to the student-university relationship, along...2. Taylor, Justin Q., et al. “Avoiding a lost generation of scientists.” eLife 5 (2016): e17393. Pubmed...

...facilitate effective teamwork, manage day to day operations within their workplace, and pursue leadership...need regular re-direction and alignment with the overall mission of the NPA. You must accommodate many different...learnt to try and bring a group of people to an overall consensus and developed negotiation skills that...would be useful in many careers. I learnt that the overall voice may not always be perfectly aligned with ...to think about their career options, gaining transferable skills such as leadership skills can only assist...

...the spotlight — Cas3. Cas3 can induce genomic alterations on a scale that other Cas effectors can’t, while...maintaining relatively high efficiency. These alterations are achieved through DNA degradation, thanks ...that the cell can use to repair. Figure 3: General overview of the Cascade-Cas3 mechanism. Created... gRNA. Cas3 appears to have fewer off-target alterations, likely due to the multi-Cas mechanism. These...has the potential to be incredibly useful in therapeutics. Imagine deleting a problematic copy of a gene...

...controversy, and the interactions of organic molecules like proteins with magnetic fields generally appears to ...magnetic responses: EGFP, mScarlet, and AsLOV2. After several rounds of semi-random mutagenesis and screening...behavior has already been successfully reproduced by several other labs. Andrew and Maria expect that further-improved...tools.  Embracing science in the open The York Lab operates with a strong commitment to exploration and collaboration...

...be able to tell you the laser configuration. In general, if two fluorescent markers are read by the same...to become a perpendicular vertical line.     In general, most spillover between samples can be fixed with...anyone just starting to use flow cytometry, it’s generally recommended to start compensating with single-... compensation panels below each plot.  The general rules of compensation apply to your experimental...research background in CD8+ T cells and cancer immunotherapies. She is currently a research technician in ...

...how best to prepare. Maybe you’re a presentation veteran, but have difficulty answering questions. Or perhaps... virus of interest causes cancer, as well as therapeutic strategies to target it, than you typically would...topics,” so be sure to be well-read on the recent literature in your field. Take your time: We often get so...making it difficult to see the actual poster or interact with the presenter. If you don’t have time to ...at the University of Arizona. She studies the interaction between Human Papillomavirus and the innate immune...

...light generated by sunlight to observe single gold particles with diameters less than 4nm. Lateral view...field-of-views (6). For more comprehensive reviews on several other LSFM configurations’ various features see...microscopes are less efficient than the modern cameras used in LSFM. In LSFM the laser light sheet, typically...microscopy . Since image acquisition is performed using cameras with electron multiplying couple-charged device...

...paCas9-1 using nMagC714 instead of nMagHigh1-C714, generating paCas9-2. This change did not significantly alter... alter the system’s efficiency at generating mutations when activated with light and lowered background...out the Cas9 domains in their split fusions and generate a photoactivatable nickase and a photoactivatable...protein due to being fused with high-affinity interacting proteins Coh2 and DocS, respectively. The use... the versatility of this approach. This study generated the first optically active saCas9 protein. Stop...Charpentier E, Möglich A (2013) Engineering of temperature- and light-switchable Cas9 variants. Nucleic ...

...formation on agar plates, less stable at high temperatures and in acidic environments.   Carbenicillin   ...mechanism. How to choose between the two? Carb is generally a ‘better’ antibiotic than amp: it is more stable...inactivates it. The NPTII gene confers resistance to several aminoglycosides, including G418 which is commonly...induce cell death by intercalating into DNA and generating DNA breaks. The Sh ble gene provides zeocin resistance...., & Brooks, G. F. (1986). Effect of storage temperature and pH on the stability of antimicrobial agents...

...Brunello library consists of 77,441 sgRNAs with an average of 4 sgRNAs per gene and 1,000 non-coding control...identifying strong hits in genetic screens, they do have several limitations. CRISPRko results in a complete loss... and the ORF overexpression library identified several overlapping genes, most genes were unique to the...Applications for the Root and Doench CRISPR libraries Overall the Root and Doench lab’s have proven their optimized..., Kendall R., et al. "Up, down, and out: next generation libraries for genome-wide CRISPRa, CRISPRi, and...

...hundreds of samples, with an average of 10 plasmids per deposit. Each week an average of 16 deposits is received...Laboratory Operations Manager at Addgene.  In addition to helping with daily laboratory operations, she also...stored right away. Every week, the lab performs an average of 140 transformations. The lab typically transforms...