...journal club! What happens during journal club? There are many common ways to run the journal club. Some...competitive and relevant. Enter: the journal club.  A journal club is a recurring meeting amongst scientists...meetings. More recently, online journal clubs have become popular, including those based on Twitter. At my former...the facilitator(s) of the journal club? Where will the journal club take place? Preparing for the first...Before the first journal club, think about the meeting itself A journal club should aim to have many of...should almost universally include the questions “Would you attend this journal club again?” and “How would...of the journal club?” You also likely want to evaluate the “success” of the journal club. This can be a...

..., Musicophilia includes case studies of relevant neurological atypicalities, including a patient with ...Addgene's book club Indeed, reading is ingrained into Addgene’s culture. We have a book club in the office...From Addgene Book Club to our on-site book swap/library, reading is part of the culture here at Addgene...protagonist, some clever magic, and a satisfying conclusion? In this series, there is always another secret...in 2011 and is still going on today! Addgene Book Club (ABC) founder, Eric Perkins, is Addgene’s Director...explains the origins of ABC: I founded the Addgene Book Club (aka ABC) in 2011 after having numerous conversations...book-loving Addgenies. I’d never participated in a book club before, but I had a few reasons for launching this...

...up cell clumps Success in a flow cytometry experiment depends on a variety of factors including sample ...instead of vortexing. Include proper controls To ensure accurate data analysis, include a complete set of ... by denaturing proteins and dissolving lipids including those of the cell membrane. In some cases, this...whole blood sample will contain a mix of cells including granulocytes, lymphocytes, and monocytes. Due ...different populations of immune cells express unique Clusters of Differentiation cell surface markers (CD) these... through a nylon mesh cell strainer to break up clumps before running on the instrument. To sustain cell...staining.  If you’re using antibodies for detection, include an unstained control with cells that went through...

...optogenetics toolbox with CRY2clust Article contributed by Brook Pyhtila  Listen to the CRY2clust podcast segment...peptide. CRY2clust should prove a useful tool to further probe cellular function. The CRY2clust Plasmid...proteins in response to blue light. This tool, CRY2clust, was created by adding a 9-residue peptide to ...CRY2, although some studies have shown that this clustering only happens under certain conditions. With...With its additional 9 amino acid peptide, the CRY2clust domain enables fast and reversible oligomerization...reversible upon termination of light stimulation. CRY2clust enables faster assembly and disassembly dynamics...sensitivity to light, and better distribution of clusters without accumulation in subcellular compartments...

... antibodies, include controls where each primary is omitted but all secondaries included to uncover secondary... shifts indicative of tau phosphorylation. By including the positive control, you will be able to confirm... addition to experimental samples (?). B, The inclusion of biological replicates such as multiple mice...Replicates can be technical or biological and are included to improve the precision and accuracy and therefore...repeated measurements of the same sample and are included to assess variability in the experimental protocol...: Microscopy-based antibody experiments should include a primary antibody specificity control, secondary...antibody. The secondary antibody control should be included with each experimental run (Burry 2011). Autofluorescence...

...particular interest this fall:  The BioBuilderClub BioBuilderClub engages high school teams around the...runoff from winter icing.BioBuilderClub is the only informal synthetic biology club of its kind, running...classes, some pick and choose a few lessons to include each year; others use BioBuilder as an afterschool... work with one of five BioBuilder kits — which include lab materials and reagents such as droppers, centrifuge...biology.Launching with a kickoff meeting in October, BioBuilderClub supports teams through monthly online group...completion, from pitch to prototype. Last year’s BioBuilderClub teams’ biological engineering designs and ideas... year from remote scientist mentors and the BioBuilderClub community. Registration is open through October...

...them. Other popular reasons to march include: General inclusion of science to inform policy (17). The...6). Many of the new administration’s actions, including taking down the climate change page on whitehouse.gov... consequences of a march. Three of these risks include: 1) Lost trust Perhaps the greatest risk is that...with one unifying interest (the advancement and inclusion of science) they come from many different places...see and understand data themselves can come to conclusions themselves instead of depending on scientists...that ‘facts’ are subjective or alternative. Inclusion of data informing policy specifically related ...the most profound impact (15). Diversity and inclusion in science, technology, engineering and mathematics...

... the Chi-Bin Chien lab (Kwan et al., 2007) and includes four promoters, six fluorophores with nonoverlapping...promoters for cell types directly linked to disease, including motor neurons (-3mnx1/Hb9), pan neuronal (elavl3...CyOFP1), some of the exciting properties of mCyRFP1 include a redshifted-emission spectrum compared to that...The set of CyRFP constructs available at Addgene includes bacterial and mammalian mCyRFP1 or CyRFP1 expression... was initially limited by off-target effects, including the conversion of other cytosines near the target...knockouts replace entire protein-coding regions (excluding start and stop codons) with kanamycin cassettes... cassette via Cre recombination. Each knockout includes unique barcodes and universal priming sites to...

...you have many career options available to you including careers in academia, industry, non-profit, and... to maintaining a status in the United States including green cards that are well-tailored to meet the...specialized knowledge in a field of human endeavor including but not limited to biotechnology, chemistry, architecture...publications, and plan for future activities; including the interest of the greater scientific field, ...entities or individuals. A successful NIW case should include evidence such as proof of US government grants ...fact is best proved through objective criteria including awards, media attention, citations to your work... discuss your work. Participating as a judge (including as a peer reviewer for journals, acting as a journal...

... an array of post-translational modifications including protein folding, formation of disulfide bridges...for protein production and purification. These include mammalian, insect, bacterial, plant, yeast and ...summarize some of the more common expression systems including their advantages and caveats to keep in mind before...Some drawbacks of insect cell expression systems include that virus production can be quite time consuming...produced by bacteria become insoluble, forming inclusion bodies that are difficult to extract without harsh...requirements as mammalian cell expression systems, including the majority of complex post-translational modifications...been used to express a wide range of proteins including antibodies and pharmaceuticals, specifically interleukins...

...2022 AAV.MaCPNS2 PNS, including ENS (mice) PNS (rats) CNS; PNS, including ENS (adult marmosets, infant...vector, which determine the vector’s tropism, including its ability to cross barriers like the BBB.)  ...application is dependent on a number of factors, including species, strain, age, sex, area of interest, and... with unique tropisms are now available. These include variants with broad CNS or PNS transduction, cell...expression control. (c) Viral delivery methods include focused ultrasound (FUS), intracorporeal (IC), ...expression or activity can be achieved using switches, including riboswitches and optogenetic and chemogenetic ...more effective than AAV9 in many other strains, including FVB/NCrl, 129S1/SvImJ, DBA/2J, and C57BL/6J, the...

...adding a triple nuclear localization signal or including fusions with Histone 2A or 2B can result in bright...however the nanobody proteins tend to get stuck in inclusion bodies. Takayuki Iwaki, Kimiko Hara, and Kazuo...described a simple method which alleviates the inclusion body problem and makes nanobody production in ...inducible promoter. The gRNA piggyBac plasmids include a mRFP reporter. With this system, they could quantify...Addgene The MoClo CRISPR/Cas Toolkit for plants includes 95 plasmids consisting of CRISPR/Cas nucleases... uses CRISPRi to control gene expression. This includes a CRISPRi-based synthetic oscillator, bistable...of norepinephrine. These norepinephrine sensors include GRAB_NE1h (high affinity) and GRAB_NE1m (medium...

...not worried about including irrelevant events in my analysis, as these will get excluded through further...gate on single cells and exclude any events where multiple cells were clumped together. (B) Instead of...gate to identify cells, the single cell gate to exclude duplicate events, and the Live/Dead gate to focus...your cells of interest, you will likely need to exclude all duplicate events (two cells stuck together)...gated my single lymphocyte cells, I now want to exclude all dead cells from my analysis. For this measurement...Lastly, you learned which gates should always be included in your analysis, regardless of your experiment...Sharpe at Harvard Medical School. His projects include investigating negative regulators of T cells and...

.... Conclusion: Always, always, write a brief conclusion of your analysis, even if your conclusion is ...is “this approach is not ideal because…” Including a conclusion for each analysis or task will keep you... approaches to analyze large data sets (myself included) and we are faced with finding the best system...begin, briefly state the goal of the analysis, including background on other analyses that have inspired...where you also record your goal, approach, and conclusions (see above). Keep original files in a separate...

...The CRISPaint toolkit includes several donor plasmids for protein tagging, including luciferase (NanoLuc...regulatory sequences, including protein enhancers and repressors, can be included in the repair template...have been able to study many cellular processes, including mitosis, mitochondrial fission/fusion, nuclear...accepts the homology arm repair template. SapTrap includes a prebuilt donor plasmid library containing several... tagging genomic loci in other model systems, including mammalian cells. CRISPaint (CRISPR-assisted ..., thanks to the modular arrangement of vectors included in tagging systems like SapTrap, scientists will...

...(Figure 4), our catalog includes vectors in a variety of serotypes, including the retrograde AAVrg and...and resources. As editor of Addgene’s eBooks — including the recently released Viral Vectors 101 2nd edition...system can be affected by any number of factors, including the age of the model organism, injection site,...resources for producing vectors from plasmids — including the protocols we use in our lab — being able to...engineered ligands (Figure 2). Biosensors, which include calcium and voltage sensors, are the broadest category... for vectors in the “controls” category, which includes things like viral vectors that deliver GFP. ...community. Thoughtfully prioritizing which preps to include in our service allows us to make ready-to-use viral...

...choose, it is essential to include proper controls. As a positive control, include a sample that is known ...working and should always be included.  For a negative control, include a sample that does not contain... the target!) and use them in numerous fields including research and development, diagnostics, drug discovery...background staining. Antigen-containing samples often include undesirable molecules that may bind non-specifically...specific than other methods. In addition, the assay includes both primary and secondary incubation steps, increasing... protein, especially one with tags, be sure to include a sample that expresses the endogenous protein,...

... identifying lineage markers, like many of the cluster of differentiation (CD) proteins found on immune...single-fluorophore-conjugated antibodies. These methods can include labeling markers with multiple proteins/antibodies...fluorescence-activated cell sorting (FACS) enables you to include further (non-lineage) markers alongside, e.g., ...unlabeled cells of interest while the rest can be excluded, or “dumped”. From left to right: B cell, CD4+...as proteins of the secretory pathway. Those can include cytokines and chemokines; enzymes like kinases,...solution can be the same). Examples of such targets include transcription factors, histones, and DNA repair...still be easily gated on during the analysis. Conclusion Congratulations! You made it through this blog...

...Whenever possible, include positive and negative controls. Positive controls typically include cell lines or...several variations of the protein transfer step including wet, semi-dry, and dry but all follow the same... Figure 2: The western blot dry apparatus includes a stack of filter paper, a copper anode/iblot ...data sheet thoroughly. Vendors will frequently include suggestions for working concentrations, blocking...blocking buffers, and incubation times. If the vendor includes publications that cited the antibody, consider...It is important to note, however, that you must include a loading control when assessing protein abundance...western blot. When planning a western, take care to include the proper positive and negative controls, choose...

... software. In fact, SnapGene’s feature library includes nearly 3x as many features as could be detected...also eliminates many of the extraneous ORFs that cluttered our old plasmid maps. Clearly Annotated Features...greater than our previous mapping software and now includes Type IIS restriction enzymes that are commonly...tooltip information and sequence analysis tabs including: Maps This is the first tab you’ll land on...enzymes that cut the given sequence. The table includes enzyme names, the locations where the enzymes ...features detected in the plasmid sequence. The table includes feature names, locations, sizes, colors used to...detected in a given nucleotide sequence. The table includes primer names, sequences, binding site locations...