...the CRISPR toolbox. We'll cover how Cas13a was identified, the structure and function of Cas13a with a ...cleaving CRISPR nuclease Cas13a was originally identified in 2015 (Shmakov et al., 2015). They were using...the bacterial genome. From this analysis, they identified 53 potential candidate genes that fell into 3...recognized and cleaved its target RNA sequence as specified by the crRNA sequence, it adopts an enzymatically...nucleotides containing a sequence of interest (red) is amplified using Recombinase Polymerase Amplification (RPA...Reverse-transcription RPA (RT-RPA), respectively. Amplified nucleotides are combined with Cas 13a in complex...enzymes continue to make a splash in the CRISPR field. We feel that the culture of sharing within the ...

...completing the research will advance the field, create a new field, impact disease treatment, etc., again...that search committee members are experts in your field. Not true. Even if you are applying to a department...possibly the Dean, and even some people in your field that still need to understand your specific questions...display your deep knowledge of your experiments and field. It's much more impressive to answer an expert's...DO NOT need to present a detailed review of your field or the finer points of your techniques. You DO need...need to be prepared to know the literature of your field and all of the caveats and alternatives for an approach...correct positions. What is the knowledge gap in the field? How will your approach resolve this problem? (2...

...approximately one-third of cellular proteins are modified through phosphorylation (1). However, the expansion...databases of post-translational modifications, identifies a staggering ~250,000 phosphorylation events ...necessary for study. Moreover, the ability to isolate purified phosphorylated products usually depends on tools... vitro. The establishment of a system in which purified, recombinant protein could be produced with site-specific...necessary for phosphoserine incorporation is a modified bacterial strain, C321.ΔA. Because the phosphorserine...the TAG amber stop codon, using the OTS in an unmodified bacterial strain would result in read-through... of interest. Two expression systems have been verified for use by the Rinehart lab: pCRT7 NT TOPO E17TAG...

...1: Guidelines for the data fields in the deposit spreadsheet Data Field Description Plasmid ... plasmid and will help a scientist outside your field understand if this plasmid will work for them. ...synthetic biology, affinity reagent/antibody, unspecified, or other. If other, indicate type in “Vector...other, indicate strain in the “Growth Strain Other” field, and contact Addgene. We may require that you send...explanation in the adjacent Patent/License Explanation field. Filling out and sending the deposit spreadsheet... and a pop-up instruction box for the “Purpose” field. If you ever have a question about what should go...

... structure of proteins. Most proteins are then modified via an array of post-translational modifications...interactions and functions. Proteins can also be purified for use in molecular biology research (eg. polymerases...polymerases and other enzymes might be purified and used to manipulate DNA), or in medicine (e.g. insulin...translate and modify your protein of interest. Modified proteins can then be extracted from lysed cells... interest, and then lysed. Protein can then be purified from the cellular debris. There are several popular...systems, proteins are assembled in vitro using purified components of the transcription and translation... ability to assemble proteins with labelled or modified amino acids that are useful in different downstream...

.... Once you are satisfied with the plasmid information, click on the blue "Save Verified Data and Return... to submit published or unpublished plasmid(s). Fields marked with an asterisk are required. If submitting... hundred to millions. Pooled libraries must be verified by next-generation sequencing. Please see the ... theoretical — it does not have to be entirely verified by sequencing. Upload any support files that will...saved and the page will open a new set of entry fields to describe the next insert. Repeat as needed for...Bacterial Expression vector. If your plasmid can be classified in multiple categories, please check all of the... that the material I am submitting to Addgene qualifies as BL1 or BL2 as defined by NIH guidelines. I ...

...discoveries in various biological fields. In 2015, the Fujii Lab identified non-coding RNAs associated with...adaptations of this system have been made in different fields of study. In addition to the identification of ...incubated with fragmented chromatin or library of purified DNA for purification of a specific chromatin or...Biotinylated dCas9, with bound DNA, can then be purified by streptavidin high-affinity purification. This... malignancies. Additionally, the Tapscott Lab identified regulators of DUX4, a transcriptional regulator...CRISPR Cas9-guided chromatin immunoprecipitation identifies miR483 as an epigenetic modulator of IGF2 imprinting...

.... Figure 1: Silver staining of purified and non-purified AAV subjected to gel electrophoresis. ...adenoviral helper sequences. AAV preparations are purified by iodixanol gradient ultracentrifugation and ...and an internal control of known titer (protocol modified from Lock et al., 2014.) Previous experience suggests...within representative vector preparations was quantified with electron microscopy after negative staining...staining. Empty vector particles can be identified after negative staining and appear darker than full vector...droplet digital PCR (ddPCR) titration protocol (modified from Wang et al., 2018). Briefly, 293T cells are...

...you stand out.  In some fields, a cover letter is vital, particularly in fields where communication and...and writing are key skills. Other fields, often highly technical or skills-based ones, may not use them...process, it's a good idea to talk to people in the field to understand what the norms are. What is a cover.... They can be skills-based: if you're switching fields, you can use your cover letter to explain why your...scientist with over [number] years of experience in [field of research], and have excellent [list the most ...that stage, cover letters can be a barrier for qualified applicants. In short, while a cover letter can...

...previously identified essential genes while previous Cas9 and shRNA library screens only identified  60% of...a 10% false discovery rate and identified 67% of previously identified ricin toxicity genes. The screen...non-essential genes for both pathways had previously been identified (Hart et al, Bassik et al). The library performed...essential for growth and would not be expected to be identified in a CRISPR knockout screen. Improving the Design...difference in depletion of ricin sensitizer genes identified with short and long guides. Together these results...

...studies. Additionally, Spinach, the first aptamer identified to bind DFHBI, is highly unstable in vivo due...site-directed mutagenesis of Spinach. In doing so, they identified the nucleotides that contribute to the instability...Broccoli, the other green fluorescent aptamer, was identified through directed evolution. First, a pool of ... that activate the fluorescence of DFHBI were identified using systematic evolution of ligands by exponential...hydroxybenzylidene-imidazolinone-2-oxime (DFHO), a fluorophore modified from DsRed. DFHO shares many structural properties...Mango, coined Mango II, III, and IV, were then identified through microfluidic-assisted in vitro compartmentalisation...possibility for scientific advancement, and in a field with as much potential for growth as RNA biology...

...other scientists within their fields than scientists outside their fields Figure 6: Follower Network of... follow scientists within their field rather than outside their field. Lower: Follower networks of subgroups...used in this post were created using data from or modified from Ke et al. 2017. 1. Among scientists, historians...Scientific Disciplines on Twitter. Twitter users were identified as scientists if they had a scientific title ...

...all you need are some of your favorite proteins purified to carry out your plan. With a little thoughtful...binding protein (MBP) is a 45 kDa tag which can be purified (along with its fusion protein) with amylose resin... prevents insoluble fusion proteins from being purified by GST (maybe try MBP if you are having solubility...Copper, etc.), allowing His fusion proteins to be purified by single step metal affinity chromatography. ...stringent washing. Nonetheless, this tag can be purified with several different resin types and is a fairly...Coligan, J. E., Dunn, B. M., Speicher, D. W., & Wingfield, P. T. (Eds.). (2001). Current Protocols in Protein...

... let’s say you identified gene target X for BRCA-deficient breast cancer and identified several candidate...The biomedical field is often concerned with understanding the cause of diseases and how to treat those...requires drugs. While we often think of these two fields as separate, they are deeply intertwined. In this...to their target, or their direct target can be identified if it isn’t known. If a strong candidate emerges... exciting time to be in the biomedical research field! References and Resources References Fellmann, ...

...about science communication, best practices in the field, and even getting into some active science communication...laypeople.  Laypeople are simply non-experts in the field of science you are communicating from. For instance...communication by and to scientists in (roughly) the same field, often through manuscripts, seminars, or posters...to engage students at all levels.      Jargon is field-specific terminology or language. This can refer...refer both to terms that are only used in the field and terminology that is widely used but has a very specific...specific use-case (definition) in a specific field.  For example, while “PCR test” has recently transitioned...

...outside your particular field of study Assess your interest in a different field Outreach activities...Individual Development Plan (IDP) and uses it to match qualified scientists to jobs in industry, academia, and ...brainer! Engaging audiences from across the hall, fields, and other professions increases your chances of... Facebook, etc.) to learn about available jobs, fields, and trends “Never Leave an Empty Seat” at conferences...Communication Networking As the number of qualified scientists entering the workforce increases, EAs...

...plants can be easily identified and have readily observable phenotypes quantified. Fluorescent reporter-based...However, it is important to note that naked and unmodified dsRNA elicits immunogenic reactions that complicate... as the starting material (DNA-based vector or purified RNA) and the initial state of the dsRNA upon activating...target required physical isolation of the visually identified subpopulation. For traditional cell culture, ... Instead, microscopic plants must be visually identified as undergoing RNAi through the fluorescent reporter...Artificial MicroRNAs for Multiplex Gene Silencing and Simplified Transgenic Screen. Plant Physiol 178:989–1001...

...products from both. These products are then purified, quantified, and sent for sequencing. Analyzing your...exciting and versatile offerings! We recently re-amplified our distribution stock of the Brunello Human gRNA...taken to ensure that all of these plasmids are amplified and none are lost. Each Addgene library has its...are differently sized, smaller plasmids may be amplified preferentially, skewing the composition of the... overrepresented plasmids (plasmids that were amplified preferentially). You can also rank the plasmids...

... to look to the United Kingdom’s Nuffield Council. While the Nuffield Council has recently published a.... While this claim has yet to be independently verified, the watching world erupted in controversy. Briefly...mitochondrial diseases. The key success of the Nuffield Council’s report is that when it became clear ...the transmission of mitochondrial disease;” The Nuffield Council began their research into both novel and...any other germline editing technique—too. As the Nuffield Council emphasized, the key to moving research...

...audiences in other fields, like when collaborating with colleagues in other fields or writing grant proposals... to draw attention and support to work in their field. It can be hugely beneficial to pose these why questions...public talk, ask a friend who works in a different field to listen to you practice.  Ask them what they learned...main goals: To help graduate students in all STEM fields become excellent science communicators. To empower...students to become ambassadors for their respective fields. To help attendees launch new organizations that...