...interaction allows scientists to easily purify and identify any proteins that are tagged with biotin using...biotin-phenoxyl radical in the presence of hydrogen peroxide. This radical reacts with electron-rich amino ...hours like BioID. APEX has allowed scientists to identify the proteomic map of the human mitochondrial matrix...multiple rounds of directed evolution until they identified a mutant with significantly increased sensitivity... binary protein-protein interaction as well as identify additional interacting proteins in one simple ...uses proximity protein labeling based on BioID to identify RNA-protein interactions (Ramanathan et al., 2018...monoglycine residue at the N terminus with a small peptide (LPETG). In EXCELL, the mgSrtA variant can be displayed...

...the gRNA (composed of a ~20 nucleotide targeting sequence and ~75 nucleotide scaffold sequence) is consistent... storm owing to its ease of use and utility in a wide variety of organisms.  While much of current CRISPR...being realized. There are a variety of reasons to consider using genome editing to change the genetic code...developed in other model organisms (see our CRISPR guide for common CRISPR principles). However, plant-specific...hereafter referred to as Cas9) and a single stranded guide RNA (gRNA) is sufficient to modify the genome of...nanostructures (Zhang et al., 2019) or cell-penetrating peptides (Santana et al., 2020) and plants viruses (barley...deliver CRISPR-Cas9 to plant cells and should be considered alternative to PEG or agrobacterium-mediated ...

... you can feel lost. Use this post to explore the wide range of careers available to scientists and open...from Grad Student to Entrepreneur Science careers divided by the most pertinent skills required Research ...and carry out in-depth research is applicable to a wide range of industries and is a highly desirable quality...including: Science Reporter Scientific Sales Epidemiologist Scientific Recruitment Meteorologist Research...explore the different options available and then decide on a course of action. Instead of rushing in, you...into a career, do your research Once you have identified possible careers, the next step is to research...computer programming skills? Other things to consider are wage, opportunity to advance, and how many...

...team of twenty excited undergrads. Team IISER-B’s idea for the iGEM 2019 is to take bacteria out of their...objective is to induce tolerance towards cold, we decided to focus on a special class of enzymes expressed...usually defined between a fairly nippy 20°C to a decidedly uncomfortable 45°C). Their more adventurous cousins...fruit juice clarification, meat tendering, and oligosaccharide synthesis. A major incentive to use cold-active...industry as it can potentially help minimize the incidental losses like spoilage and improve the productive...inefficiently that no industrial operation would ever consider them worth the trouble.  Enter ELSA: E. coli Learning...Acclimatisation To resolve this impasse, our team decided to create a hybrid system where we could incorporate...

...’s social media toolbox. Besides advice for using Twitter, this post provides suggestions for using LinkedIn...Now I write for the @Addgene blog & I'm my lab's resident #scicomm expert. What tips do you have for using...discuss how scientists can start using Twitter and provide real-life examples of how they and others have ... February 13, 2014 2. "The Scientist's Top 10 Guide to Twitter" blog post from Dr. Paul Knoepfler Hopefully...pknoepfle 3. "Social Media as a Scientist: A Very Quick Guide" from Nature Jobs Blog While Twitter is a popular...sheet from Science Media Savvy This tip sheet provides advice for tweeting and blogging as a scientist...Addgene blog. 5. "Social Media in Your Working Day" video from Science Media Savvy At this point, you might...

...experimental needs, and can transduce (infect) a wide range of cell types with a high efficiency. However... part of the host genome that can replicate and divide along with the endogenous DNA. AAV also has low...AAV is also a cool tool in the laboratory! It is ideal for whole animal administration, such as in mouse...alternative virus, or other strategies may need to be considered if size constraints are an issue. Adenovirus...hamsters, although no cancer-connection has been identified in humans to date. Over fifty serotypes of human-infecting...human-infecting adenovirus have been identified so far! Adenovirus generally has high immunogenicity, ...viruses used in biological research, you might have an idea which ones could be helpful to you. Interested in...

.... Manufacturers rarely provide the construction plans or CAD (computer aided design) drawings of their...users the chance to understand the "black box" provided by the manufacturers and enables customized experiments...limitations into account in the development process. Widely used, commercially available tools such as the ... the community to regularly replicate designs worldwide. Distributing hardware via globally established...MIT), and documentation (e.g. CC-BY) allow users outside the project to modify the raw and design files,...of work from universities and research centers provides a potential conflict between traditional IP-based... for innovative organizations and companies to provide problem-solving platforms.     Fig. 3: ...

...” has me considering the microbiome more often than not). If you’re someone who’s considering getting ...definitely enjoyed the lab!) and would therefore consider myself a neuroscience novice (newb). In this post...entire story-telling session organized by Story Collider and which featured a talk about the different ...Developing better AI programs gives neurobiologists new ideas about how the brain functions and vice versa. Even...getting into neuroscience from the mol bio side you’re sure to have many opportunities to learn about and develop...large data sets. Whether these data sets include videos of behavior, many many images of brain regions,...particularly interested in using AAVs because they provide a highly versatile means of delivering genes to...

... between these methods is that both attempt to identify new players in a biological process. Players can...approaches can be found. As an example, say you are considering either a cellular screen or a selection. A selection...molecules/drugs. With the advent of CRISPR, genome-wide knockout screens are possible with gRNA libraries...specific to your scientific question. Things to consider when selecting an assay system include: How specific...t yield any hits. Make sure you plan ahead and consider these questions to save yourself a headache later...population. For these experiments, the goal is to identify manipulations which either enhance or suppress...scene, there are other options which are worth considering. RNAi screens are similar to CRISPR, but instead...

...maintenence within an E. coli cell, but vectors can be widely utilized across many different cell types and each...budding. Yeast Centromere plasmids (YCp): These are considered low copy vectors and incorporate part of an ARS...are most similar to bacterial plasmids and are considered “high copy”. A fragment from the 2 micron circle...most commonly used selection markers in yeast and provides the element needed to overcome the auxotrophy ...as additional uses for said element. This link provides a more extensive list of yeast auxotrophic markers...auxotrophic selection in E. coli? Additional considerations HIS3 L-hisitidine no S. cerevisiae  no yes...experiments. MET15 L-methionine and overproduces hydrosulfide ions yes - Grow with methylmercury. S. cerevisiae...

... approaches a host cell surface are complex polysaccharide structures, many of which are easily varied...proteins could function as surrogate antibodies for identifying and characterizing sugars and sugar-containing... anything else, making it difficult to quickly identify possible gene function. In this post, I will describe...assay, including examples of RBPs successfully identified using this method. Step 1: Prepare a random fragment...transformants for appropriately sized inserts. We consider a library worth using if at least 40% of colonies...overnight), flip the membrane over and place it colony-side up onto a fresh LB plate containing an appropriate...onto a lysis solution-soaked filter paper (colony-side up) and leave for 1 h at room temperature. After...

...missense variants identified from 58 individuals with GNB1 disorder since the identification of this disease...and Accelerate Research Discoveries) Challenge provides grants for rare disease research. This year is...other symptoms as discussed in this educational video. “SLC6A8 belongs to a large family of solute channels...perform a functional screen in mammalian cells to identify any solute channels that can compensate for the.... “This proof of concept study will help in identification of new targets that could greatly enhance our...keratinocytes,” says Mellett. This would help the lab identify therapeutic targets in the future. Congratulations...will express these candidate genes in TS iPSCs to identify whether any rescue cell migration and contraction...

...want to reconsider your approach, and try a different cloning strategy. Once you've identified potential...plasmid and all the steps you will have to take. Decide which technique you are going to adopt (i.e. restriction...digestion on an agarose gel. If the digestion does not provide the fragments of the size you expect, check the...region of the primer to 40-60 bp. It is always a good idea to perform primer optimization, especially if you...that, they can be open source (i.e. primer3) or provided by the major companies selling PCR reagents. Nowadays...Now that you've read our plasmid troubleshooting guide, you hopefully have some new strategies to tackle... the lab he develops new assays and processes. Outside the lab, he enjoy the outdoor in all the possible...

...secondary considerations to validation, application specificity, and experimental considerations - no other...you know which ones to try?  Read on and we will provide you with some advice on the antibody selection ...antibody first look at the application-specific data provided on the manufacturer’s website. Because of how ...has been validated in your specific application. Consider also that there may be variations within an application...section then a directly-conjugated antibody will provide better resolution as these immune complexes are...abundance on a Western blot, a secondary antibody is ideal as it will help to amplify the signal. Some applications...the species that the antibody cross-reacts with. Ideally you will find an antibody that has been validated...

...enzyme-bound hydropersulfide that is then transferred to thiol-containing acceptors. These hydropersulfides modulate...live cells. His work focuses on the enzyme hydropersulfide-propagating enzymes 3-mercaptopyruvate sulfurtransferase... susceptible to redox modification, the MPST hydrosulfide can be transferred to the roGFP2 cysteine thiols... to better understand the processes that happen inside the cell,” says Pedre Pérez. Figure 2: ...she studies opioid use disorder. Her goal is to identify cell populations in the brain that contribute ...neurons at specific firing frequencies. She hopes to identify how in vivo firing frequency of PV neurons affect...genetically encoded voltage indicators. Tian developed a video-based high-throughput screen for directed evolution...

... 1 we provide an example of such a comparison between cyan fluorescent proteins using widefield imaging... which usually are undesired properties. In the ideal situation, a fluorescent proteis should emit a stable...since different fluorescence imaging strategies use widely varying amounts of excitation power (Shaner et ... intense light (for a brief time), whereas in widefield imaging fluorophores are excited with relatively...measurements and hope this information can help guide your understanding of reported data. Photobleaching...Measuring photostability at realistic powers will provide better insight into the photostability in the intended...situation. Measuring photostability in living cells provides a much more realistic view of photostability. ...

...Instead they are detected with tyramide signal amplification. Tyramide signal amplification occurs when...are secreted from cells due to their IgGκ signal peptide sequence, making them easy to harvest from cell...when HRP converts a fluorescently labeled tyramide molecule into a highly reactive radical, which is then...Detection Method Uses Pros Cons Antigens HRP (P) Tyramide signal amplification of HRP Staining cultured...Enzymatic reactions on tissues can be cumbersome Tyramide reagents can be expensive Can only stain with ...of a nanobody that would make a useful RANbody, consider leaving a link to its sequence in the comments...immunodetection If you enjoy DIY lab projects, consider making you own DNA ladders with these plasmids...

...-art methods of sex and gender analysis and 2) provides case studies with concrete examples of how gender...capacity of stem cells. Yet, very few researchers consider the sex of their cells when doing experiments—...between male and female cells. The result of not considering the sex of the stem cells was that their male... in Norway, the team realized they should also consider the sex of the stem cells. In this case, they ... many researchers I’ve talked to who’ve never considered the sex of their cells. For one thing, cells ...hard to determine. To remedy this, Shah et al. provide a table indicating the sex of the most commonly... there are numerous methodological articles to consider—we have loaded those we are aware of in our checklist...

...proteins had to be re-engineered for each gene, making wide-scale gene activation seem next to impossible. The... systems only require changing the programmable guide RNA. Gene activation by dCas9, also referred to ...specific sequences, CRISPR/Cas systems rely on a guide RNA complementary to the sequence of interest. Upon...VP64, a strong transcriptional activation domain. Guided by dCas9, VP64 recruits transcriptional machinery...of VP64 per each dCas9, SunTag uses a repeating peptide array to fused with multiple copies of VP64. By...two-component system dependent on gRNA design (SAM) or peptide design (SunTag). This streamlines its delivery,..., Crawford GE, Reddy TE, Gersbach CA (2013) RNA-guided gene activation by CRISPR-Cas9–based transcription...

... your video call.  Expect technical difficulties. To try and circumvent this, test out the video conferencing...development, and company culture.  It’s also a good idea to prepare 1-2 unique questions for each interviewer... homework before the interview. If you are not provided with a list of who will be interviewing you, ask... your audience. Connect your prior work or a new idea to the company’s area of expertise. This has potential...difficulties. Have your presentation on a flash drive and consider bringing your own laptop as a backup, if the company...the interview, if possible. Otherwise, read the video conferencing instructions you received from HR carefully...after completing each section of your talk. Some video conferencing software allows participants to “raise...