...few minutes. The host could also set aside Breakout Rooms to divide the audience into small sessions and...To date, aside from doing science, he has assisted many researchers in gaining the confidence to launch... at the lab meeting, the team can discuss those ideas and build from those.   Figure 1: Virtual...focus on. Use collaborative cloud apps It is a good idea to use collaborative cloud tools such as Google ... and Sheets during Zoom lab meetings to chip in ideas and take minutes that can be accessed by anyone ..., where everyone stands up, disappears from the video, and prepares a coffee, or simply moves away from...I believe this is a great functionality that is widely used and could bring back a taste of that casual...

...Cas9, for which efficiency varies widely across guide RNAs, each guide tested with CasRx yielded >80% knockdown...Cas13d proteins they identified have little sequence similarity to previously identified Cas13a-c orthologs... to tap the wealth of CRISPR possibilities. To identify Cas13d, both groups used very general bioinformatic...independently process their own CRISPR arrays into guide RNAs. crRNA cleavage is retained in dCas13d and ...its low immunogenicity. But these studies also identified other advantages, including Cas13d-specific regulatory...produced very similar cleavage products even when guides were tiled across a target RNA, indicating that...Previous CRISPR exon-skipping approaches used two guide RNAs to remove a given exon from the genome, and...

...ongoing Plasmids 101 series. Plasmids 101 will provide you with an overview of general molecular biology...plasmid with an insert? Blue-white selection is a widely used method to do just that! (To read about other...lactose analogue, IPTG (isopropyl β-D-1-thiogalactopyranoside). (To be completely accurate, IPTG binds...x-gal (5-bromo-4-chloro-3-indolyl-β-D-galacto-pyranoside) into galactose and an insoluble blue pigment...encoding this section of amino acids (called the α-peptide) to a lacZΔM15-mutant bacterial cell in trans complements...engineered a multiple cloning site (MCS) into the α-peptide (represented as an orange wedge in the figure on... your DNA (red) is cloned into this MCS, the α-peptide gets interrupted as shown in cell A, and thus will...

...relationships outside your organization. The mentor in your own workplace is able to: Help you identify specific... and become an insider faster A mentor that works elsewhere is able to: Help identify the skills you... you meet, the more potential mentors you will identify. Finding potential mentors is one of the many ...done this with people I work with and people from outside my work for almost 20 years. It is good to be a... this recent blog 5 Strategies for Finding Your Ideal Mentor. One of the best was to notify your network...family and friends know you well and might have good ideas for who you should connect with. Listen to our podcast...you will need to make a change Provide perspective on core values at another company/department Give more...

...nonprofit institution, you can order plasmids covering a wide range of fields from Addgene. What you might not...this kit have been used to purify a variety of peptides including antibiotic efflux pumps and ankyrin ...followed by leader sequences (encoding small leader peptides) downstream of which you can clone your gene of...interest. Translation of the standardized leader peptide prevents secondary structures in the mRNA encoding...from resulting in the production of a downstream peptide, the transcription terminators in this series are...plasmids from this kit include the production of a suicide switch in cyanobacteria, synthetic inverters in...biotechnologically important microalgae: intrinsic suicide switch implementation in cyanobacterium Synechocystis...

...demographics and the information is often confidential (see image below). Slide of information gathered by Postdoctoral... this very idea that postdocs are acquiring training to become professors that is provided as an explanation...transferrable to other environments. Slide from Paula Stephan’s Talk (All slides can be found here). Importantly...information does not, however, prevent us from identifying problems with the structure of the academic research... research workforce, and we are beginning to identify potential solutions. Keynote speaker Paula Stephan...masters programs to prepare scientists for positions outside of academia Require that postdoc salaries be paid...coming months and welcome guest bloggers who can provide additional advice to trainees navigating the waters...

... Addgene's resident blog editor, I could take the needed steps to continue where the guide left off - ...molecules can bind to a wide variety of targets, including proteins, peptides, amino acids, drugs, metal...I love Addgene's new Antibody Guide and not just because I helped create it. But while antibodies, as...its thorns and every affinity reagent has its downsides. These reduced-in-size options often have reduced...situations where a suitable antibody has been identified but a smaller or less immunogenic version is ...light chain regions, connected by a short linker peptide. The lack of constant region makes them less immunogenic...proteins formed by two Fab fragments bound by short peptide linkers. They are bispecific, with a lot of flexibility...

... be identified using antibodies and immunohistochemistry techniques. Thus, these proteins provide a versatile...to the plasma membrane.   Our studies provided evidence that some fluorescent proteins can be, in ...facilitate its detection. Fluorescent proteins are most widely used tags because they emit endogenous fluorescence...versatile method to identify the expression of transgenes of interest. Furthermore, fluorophore variants...red fluorescent proteins) allow simultaneous identification of more than one protein of interest. However...were still cytoplasmic. Use of ‘small’ tags to identify proteins Given the limited expression of hM4Di‐mCherry...proteins when the fluorescence is not essential to identify the expression of the opsin or chemogenetic receptor...

... BioID platform for identifying protein-protein interactions, but RaPID identifies RNA-protein interactions...ligase BirA* from BioID and allows a researcher to identify proteins that bind an RNA motif of interest in... like cross-linking immunoprecipitation (CLIP) identify RNA-protein interactions by cross-linking RNAs...using proteins as bait has limited utility for identifying novel RNA-binding proteins that interact with...RNA of interest. Current labeling methods are not ideal for in vivo work. BirA* requires 16-18 hours to ...Repository for Affinity Purification (CRAPome), to identify highly biotinylated background proteins reported... not well characterized RNA motifs. This bait identified a new interaction: binding of the RC2H1 protein...

...experimental design considerations used in all behavioral studies. General considerations for behavioral ...should be avoided as this will also increase stress levels and affect behavior. A fact to consider: Rodents...behavioral testing As a general consideration for behavioral testing in a laboratory, a high level of experimental...fluorescent light bulbs, for example) that are outside the humans’ audible spectrum. The experimental ...gene expression in a specific brain area, avoiding side effects from such modification in other areas. An... particles are smaller and may be preferable if wider diffusion is required. On the other hand, lentiviral...targeted to a specific area. Some other technical considerations for stereotaxic injections are: Vector titer...

...go-to providers of materials needed for optogenetics. There are several important considerations with ...optogentics to answer a research question. The flip side of the optogenetics coin is materials science-based...dental cement (a similar procedure as implanting a guide cannula). A fiber optic patch cable is then connected...materials required for each [1, 2]. This protocol [3] provides, in exquisite detail, the various steps required...able to penetrate brain tissue a short distance so ideal implant placement should be about 0.4-0.5mm above...stimulation. The scatter effect is also an important consideration when preparing the implant itself, which brings...least 85% of the initial laser output. The same considerations apply to the polishing and preparation of the...

...with the guide RNA.   Non-target strand The DNA strand that does not base pair with the guide RNA. Despite...hypoxanthine (shown) is the nucleobase component of the nucleoside inosine. Created with BioRender.com. Although...ABEs are base transition editors. They adjust the side groups of DNA bases and leave the core ring structure...some cases, the same repair pathways that were considered problematic in the development of CBEs and ABEs...structure. In order to bind DNA, Cas9 requires (1) a guide RNA, (2) a complementary DNA sequence, and (3) a...strands called an R-loop (Figure 2). This allows the guide RNA to base pair with the DNA target strand, leaving...non-target strand unpaired and accessible on the outside of the enzyme. Figure 2: CRISPR R-loop. ...

...knockdown, or other factors, consider using one of the alternative, albeit less ideal, verification approaches...the results across runs. If the results vary considerably, then the high degree of variability in the ...replicates are distinct samples that are treated identically, used to assess naturally occurring random biological...assays have their own unique controls that must be considered (Burry 2011). Perhaps the most critical are those...wild-type and knockout samples are treated with the identical protocols that the experimental samples undergo...with its antigen or a small epitope-containing peptide to block the antigen-binding site. The sample is...detectable, it is likely due to binding interactions outside of the antigen-binding site.      Secondary antibody...

...industry, nonprofits, and publishing. The livestream video from the event can be found below along with a description...reproducibility “crisis” on our hands and instead provide a brief conceptual framework for the causes of ...protocols and use one another’s reagents with the confidence that they aren’t simply doing an experiment wrong...Luckily, many organizations have been working to provide tools that make it easier to share in-depth information...working with a plasmid from Addgene, you can be confident that you’re using the actual tool that you ordered...research needs. Other companies like NEB and ATCC can provide you with additional validated reagents, and, with...Protocols.io, JoVe, and Bio-protocol all work to provide more descriptive and interactive protocols that...

...designed small RNA, or guide RNA (gRNA). The gRNA conventionally contains a 20-nucleotide sequence that is ...Activators: A Practical Guide Additional Resources on Addgene.org Check Out Our CRISPR Guide Pages Find Validated...nuclease-null—in order to effect different perturbations. Consider, for instance, the challenge of attempting to ...interacts with a distinct gRNA that allows users to decide which Cas9 protein is directed to which target ... truncated such that it exhibits 15 or fewer nucleotides of complementarity to a target site, Cas9’s DNA...truncated gRNAs Importantly, the use of truncated guides has largely been demonstrated to result in decreased.... An additional benefit of the use of truncated guides is that they can be delivered into systems that...

... about my interests and my ideas all the time. Once, I had this crazy idea that I needed help with: wouldn... you make the most of the opportunities you’re provided throughout your PhD program.   Find people who.... They didn’t always know the best direction to guide me because they had no experience with my career... open mind and open ears. Everyone I talked to provided new data. Every new experience taught me something...troubleshooting.” Trying out new experiments and new ideas is nothing new to a scientist. Framing my efforts...efforts in these comfortable terms helped guide me through the process. As I mentioned, my graduate office...disappointed in the lack of participation, but I decided to focus on what the experience taught me. The ...

...as well as highlight some considerations for their use. Then, we’ll provide information about commonly...decision tree can help guide your choice on which systemic capsid to use. Factors to consider include the delivery...goes beyond such restrictions, you may want to consider a systemic capsid for your AAV.   Systemic capsids... definition of systemic capsids. After some consideration, we offer this definition: A systemic capsid... after intravenous administration, distributing widely throughout the organism to target large or diffuse...commonly used and emerging capsids to help you decide which ones may be suitable for your experiment. Like all...and/or its intermediate engineered precursor. Considering systemic capsids for your research? While systemic...

...disciplines. Cast your net wider, explore tangential subjects, and let curiosity guide your intellectual journey...thus feel grateful) and identify areas in need of improvement. These unrelated ideas might hold the key to...simple steps to welcome these unforeseen moments. Widening your search The first step towards inviting serendipity...ultimately enhancing your project's potential. Widening your search creates fertile ground for serendipitous...encounters that could spark unforeseen connections and ideas. Maintaining openness and looseness of spirit The... exploration of tangents or seemingly unrelated ideas. It's beneficial to carry a notebook at all times...confines of a predefined path, opening doors to novel ideas and approaches. Moreover, the ability to embrace...

...develop their career skills for positions both inside and outside academia. Inspiring future scientists At ... ever heard of a Freshers Fair? If you’re from outside of Europe, the answer is probably no, but these...be used as welcome events for new students that provide ways for students to network, find groups with ...come. Some Freshers Fairs can include fairground rides and other fun activities and even invite members...thought we’d take some time to introduce you to the idea of the Freshers Fair and some of the great biological...students so that they can help support each other and provide advice. The society organizes numerous recreational... high schools to promote chemistry degrees and provide advice to students who want to study chemistry....

...mentoring for instructors, and provides opportunities for students to exchange ideas and results with peers and...al., 2020). Many have interpreted this trend as evidence that the best teachers and mentors pass on more... Biochemistry at the University of San Diego), decided to take a different approach. His own most impactful...experiences happened in the lab, and he wanted to provide the same kinds of opportunities for his students...but one that left him overwhelmed. Therefore, he decided to adopt a more focused format, where all students...analysis, structural prediction, and bench work to identify regions of MDH that interact with citrate synthase...Meeting. Some of my students published their work identifying new genes required for stem and progenitor cell...