...publications—the pressure to produce is the same You get to interact with scientists just like you What is different...structures are common Small integrated teams, little hierarchy Real diversity and acceptance can be an issue...requirements for hiring more common Likelier to hire on general talent and not specific skills <- Can move between...where you want to go will help you focus on the transferable skills that you will need to get there. Additional...

...developed in David Liu’s lab in 2019, has added considerably more precision and flexibility to the CRISPR... repair favoring the edited strand. The first iteration of prime editor simply fused the wild-type Moloney...pairs for prime editing. Since PAM sites occur on average every ~8 base pairs on either DNA strand, many ...by double strand breaks has been widely used to generate precise changes. However, the efficiency of Cas9...system PE2, which had prime editing efficiencies on average 2.3- to 5.1-fold (though up to 45-fold) higher ...

...cell survival or proliferation, an additional challenge is that only partial or ephemeral knockdown will...from the manufacturer and/or in the published literature. Note that the antibody being published doesn... used in combination with other approaches. If several cell lines express the target at different levels...contains information about the labeling pattern of several cell lines using these two antibodies. This is ...

...Integration. Molecular therapy : the journal of the American Society of Gene Therapy. 2008;16(7):1189-1199...gene expression in a target cell. Practical considerations If you have high quality AAV, you can address...be to prepare them for the arduous process of generating viral particles. Limit tube transfer - the fewer...

...visualizing genomic loci in living cells. Previous iterations of genomic loci visualization tools using CRISPR...PP7, or boxB sequences inserted into the sgRNA generates blue, green, or red colors for labeling. A combination... PP7, or boxB sequences in the same sgRNA will generate additional colors. Image from Ma et al., 2016 ... tetraloop and went through subsequent design iterations to increase stability and minimize misfolding...

...plasmid: name, description, how the plasmid was generated, validated sequence information, and more. The...persistent identification of research reagents, several journals or organizations now recommend or require...authors to use specific PID formats. For example, several journals like eLife, Nature, and Cell use  Research...copy and paste the text attributing the PI who generated the plasmid, followed by the PID(s) of your choice...

...behind Gram’s crystal violet stain and the ion interactions that cause the stain retention in gram positive...When the negatively charged iodine is added, it interacts with the CV+, creating a complex within the peptidoglycan...trapping the large dye complex created by the interaction of CV+ and I- in the thick multilayered membrane...can be used in medical settings to determine the general type of bacteria present in an infection. This ...

...residues. Compared to MG, MG-2I has increased ROS generation when bound by the dL5** FAP.  The iodine-modifications...cells.  He et al also show that MG-2I-dL5** only generates small amounts of non-specific ROS when exposed... red fluorescent photosensitizer protein that generates ROS when illuminated with green light. They found...inactivation/cell ablation while having limited collateral damage to bystander cells. Are you ready to use...

...expression, localization, and protein-protein interactions. Beyond these common applications, researchers...when exposed to ultraviolet light. To do so, she generated fusion proteins in which Dronpa domains flanked...Proteins, or FLiPs.  Xin later optimized these first generation FLiPs by mutating Dronpa to lower its aggregation... upon exposure to ultraviolet light. Previous iterations of FLiPs only used N- and C-terminal dronpa fusions...

...of sample preparation for high-throughput next generation (NGS) sequencing have prevented laboratories ...each well of a 96-well plate. Then, samples from several plates are pooled together to create a single library...harder and harder to obtain balanced and sufficient coverage of all samples. The root of the normalization ...important for getting adequate sequencing depth and coverage in order to, for example, assemble hundreds of...

...depending on the primer sequence and the NEB polymerase used. Overall, while much of the information in the ...helpful, but does make it easy to skim the manual. Overall, it’s a great tool for someone starting cell culture... it had a codon list for each amino acid, but, overall AMINO can be beneficial for new researchers or ...

...distances. For instance, how much force can be generated by conformational changes in proteins? How can...can be addressed using R bodies: simple, force-generating protein machines that can extend and retract ... might be able to endow cells with new ways to interact mechanically with their environment.   Jessica...

...to interview, part of the interview process was several short talks by current faculty members and it just...it looks good, that’s a high that can last for several days.” Working with CRISPR Recently Doench has...since” and that he’s “very much in line with the overall mission of making science easier and more transparent...s] outreach scientists do to comb through the literature, to go and talk to the people who actually have...

...study microbiome interactions with Alzheimer's, I found others using CRISPR to generate new knockout mice...within the biological sciences, there’s plenty of interaction between neuroscience and other fields. For instance...us produce new educational content on the blog. Overall, there are many exciting things going on in the...

...expression requirements (Figure 1), but Cas12a has generally been underutilized in genetic screens because ...technique, researchers identified a synthetic lethal interaction between MARCH5 and WSB2 genes using Cas12a’s ...similarly to Cas9 screens in a compacted library. Overall, Cas12a’s ability to target multiple genes within...possibilities in both cell engineering and gene therapy.  Xiaoshu X, et al. Molecular Cell. 2021. https...

...post, we’ll touch on antibody storage, buffer considerations, and give just the lightest of nods towards...smaller than 10uL. You can leave them at room temperature when you’re working with them, but avoid freeze-thaw...okay, Blugene!    Buffers  You know what temperature to store your antibodies at, but it also matters...commercial kits.  However, antibodies conjugated to horseradish protein (HRP), primarily used in Western blots...

...content - the useful information on scientific literature, technical skills, and experiences from thousands... Nearly everything we write entails a primary literature search to collect up to date information on the...review process led to quick dives back into the literature to make sure absolutely everything was correct... typos, and hit publish. Making content is an iterative process. We are constantly reviewing the resources...

...yeast, are temperature sensitive screens. These involve raising or lowering the culture temperature to exacerbate...drug-containing media in which only cells which can tolerate the drug survive and everything else dies off....though. A selection can take the form of varied temperature, nutrient availability, drugs, and much more....

...interest by PCR generates mutated plasmids with no methylation (there are no methyltransferases in the PCR ...DNA. The other component of these systems, methyltransferases, methylate DNA at particular sequences to...restriction modification systems containing methyltransferases that add methyl groups to specific DNA sequences...

...to do a massive RNA-seq analysis. I had never interacted with a computer at the command line, and so was...keep track of the files that I was continuously generating.  I quickly found that there is no one way to...physical lab notebook, along with any figures generated from the analyses: Goal: Before you begin, briefly...control your commonly used scripts: If you edit a general purpose script for a single use, save the script...