...means that F(ab’)2 fragments may have overall stronger interaction with their antigens than a Fab or Fab...the protein will interact with other components of your experiment. These considerations will help you narrow...secondary antibodies you already have in your refrigerator will not recognize a Fab fragment. In order ...

... 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)...

...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...

...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...

...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 ...

...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...

...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...

...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...

...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...

...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 ...

...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...

...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...

...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...

...Administrative and Operations Coordinator at Addgene. In addition to her operational responsibilities, ... was a lot of blood, sweat, and, I’m guessing, literal tears (though none of the founders have EVER confirmed...to do, feel free to crowdsource! 2. Free food generally gets people to show up If you feel like you may... event planner)! If you have a very intricate itinerary, there is a higher likelihood that something will...

...they study (unless if you count normal human interaction for all of us studying human genes in test tubes...invited to attend. My job is to ensure that, in general, these labs all include materials sharing in their...Monarch butterfly food. They have chemical pathways generating interesting drug-like compounds especially cardenolides... heart disease. Caterpillars that eat milkweed generate cardenolides that tastes yucky (and is poisonous...Celina Juliano) This is a known stem cell and regeneration model. Little bits of hydra can grow back into...

.... Cas13a (previously referred to as C2c2), has several unique properties that further expand the CRISPR...other hand, uses only a short ~24 base crRNA that interacts with the Cas13a molecule through a uracil-rich...changes in the Cas13a molecule. Like Cas9, Cas13a tolerates single mismatches between the crRNA and target...interest (red) is amplified using Recombinase Polymerase Amplification (RPA) or Reverse-transcription ...knockdown to single gRNA expression. Although the collateral RNA cleavage response is well described in bacterial...

...can use your cover letter to explain why your transferable skills make you a good candidate for the position...experience or skills, often by pointing to your transferable skills. In the above example, if you don’t have...During my thesis work on [phenomenon X], I read several of your publications on [specific topic]. I am ...and grammatical errors. It does not need to be a literary masterpiece. For most jobs, the writing quality...professional. Always review grammar and spelling several times and ask someone else read your cover letter...

...their structural elements, oris interact with various RNA polymerases and proteins from bacterial hosts...ordonez, Serratia marcescens, Shigella spp., Vibrio cholerae, Xanthomonas campestris pv. campestris, X. campestris...Rhodobacter sphaeroides, Salmonella typhimurium, Vibrio cholerae, Xanthomonas campestris   In addition to incorporating... the broad host-range plasmid RK2. Molecular & General Genetics: MGG, 152(3), 129–135. https://doi.org...

... An overview of peptide synthesis. A ribosome interacts with mRNA and charged tRNAs to build a peptide...codon usage among different organisms. There are several codons that encode the same amino acid. Each organism... be harmful (Sauna and Kimchi-Sarfaty, 2011).  Overall, codon optimization may not always be the optimal...in vivo experiments and in biotechnology and therapeutics (Mauro and Chappell 2014).  Ultimately, it is...critical analysis of codon optimization in human therapeutics. Trends in Molecular Medicine 20:604–613  https...

...In their BioRxiv post, the authors generated and described several improved genetically encoded fluorescent...! Schematic of the evolution platform to generate TEV protease variants. Catalysis by TEV protease...markers can lead to non-specific labeling. They generated plasmids tagged with EB3 to mark the growing tips...plasmids containing different nanobodies (NB) were generated in pMAK:  pMAK461(NBαRabitb-IgG-Fc, Addgene #140699...

...plasmid-based experiments. What are control plasmids? Generally, control plasmids help to ensure that the observed...constitutively expresses a reporter protein (e.g., GFP or luciferase) that is either co-transfected with the test ...non-targeting control also assesses any effects of the general introduction of shRNA into your target cells. Positive...replicate controls: technical and biological. In general, technical replicates can be thought of as "plate...of Plasmid A in different aliquots of cells on several different days. Biological replicates control for...

...organized pipeline and accelerated the processing of our sequencing data.”  Recently, Seqera published a case...about our bioinformatics quest, which led us to Seqera Labs, a company that provides open source workflow...You can read it, and other case studies, on the Seqera website. ...

...communicator overall. Don’t Write in a Vacuum Your research papers should go through many iterations read ...Saturday Hack Day projects sought to make an interactive game that highlights just this point. As part... this, trainees need to be able to network and interact more with people who have taken a variety of career...write for a university publication, improving your general writing skills will help you in your academic publishing...