...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...

...results, conclusions, and acknowledgements sections. Some other additions you might wish to include are a...navigable and efficient; your questions, data, and conclusions should be clearly identified and interpreted ...smaller than in other poster sections, so you can include a lot more. Font size: Making your poster readable...equally-sized columns, where the introduction, methods, conclusions, and acknowledgements sections are inserted. ...the central region, and a scannable barcode is included that links readers to more information if they...background but want to add a bit more “flash,” you can include a subtle gradient (as is shown in all of our illustrations...

...collection of plasmids for coronavirus research, including a handful of plasmids that have been in the repository... plasmids used for this study at Addgene which include a C-terminal deletion in the spike protein to improve...Plasmids deposited from Alejandro Balazs’s lab includes those for generating pseudovirus and those encoding...the widely used qPCR and antigen tests.  Labs including Jennifer Doudna’s lab and Feng Zheng’s lab had...for detection have been made openly available including those from Andrea Pauli and Julius Brennecke. ...19 diagnostic enzymes. Plasmids in this toolkit include ones for expressing MMLV reverse transcriptase ...untagged and tagged versions of SARS-CoV-2 ORFs and include plasmids that are codon optimized. Browse the collection...

...intentionally excluded any libraries from the software, and my hope is to continue including other libraries...strategies beyond SpCas9, or species not currently included in Green Listed.  Provide the list of genes ...Input” box. Choose from different “Options”, including adding adapter sequences for cloning purposes.... output folder containing several files. These include: Output: A text file containing full information...information about the suggested gRNAs to use, including all information provided in the original reference library...Output_Compact: A text file that describes which genes were included, and how many gRNAs were identified for each gene... gRNA, as well as the suggested gRNA sequence including adapters. Output_UserInputParams: Contains all...

... open access publisher has required authors to include a note as to where readers may locate data supporting...information) for assessment of veracity of the conclusions drawn. The main difference with the PLOS policy...the time of manuscript submission. The rationale includes increasing the ability of reviewers (and, once...information is made available to everyone at once, including the generators. This practice doesn’t appear to...scientists are, however, privileged in having exclusive access to data and enjoy reasonable time to make...The fact that so few authors now query what to include suggests pragmatic adoption (although it might ...access to data is important, what data should be included? We should recognize that storing data in a sharable...

...data, you can even include your own data in a ReFigure. For example, you might include a confirmatory image...cofounder of ReFigure. Reading and exploration including replications and experiments resulting in “negative...also eLife, PLOS and Figshare). Some use cases include: Cataloging a reagent’s performance as in this...remain hidden from research papers as they are not included in traditional publications. By submitting your... currently supported by ReFigure beta) you can include it in a ReFigure. ReFigures are good for you and...

...sleek acronym for a real mouthful of a phrase: Clustered Regularly Interspaced Short Palindromic Repeats...chromosome encodes a tracrRNA (in some systems including Cas9), Cas proteins, and a CRISPR array. The CRISPR...is transcribed and processed into crRNAs, each including one repeat and one spacer. In bacteria, these ...DNA sequences. In laboratory settings, an sgRNA includes the crRNA and tracrRNA sequences in a “single-...RNAs” are transcribed from the CRISPR array and include a spacer and a repeat sequence. The spacer sequence...mismatches can be tolerated outside this region. Conclusion Ready for some more details? Check out the Addgene...

... allows it to be used in more E. coli strains including the popular BL21(DE3) (Li et al., 2021). This ...original pCas/pTargetF but has many other advantages including shorter plasmid curing time. pCRISPomyces for ...Streptomyces (Cobb et al., 2015). pCRISPomyces-1 includes Cas9, a tracrRNA, and a CRISPR array, while pCRISPomyces.... INTEGRATE consists of four major components including (1) a CRISPR RNA, (2) four proteins forming the...essential genes in a wide variety of bacteria including in pooled, genome-wide, formats. Interested in...proteins involved in the activation of transcription including transcription factors, phage proteins, and RNAP...Adding complexity, they found that they could also include a CRISPRi system for multiplexed modulation. CRISPRa...

...Addgene is building our antibody resource, which includes recombinant antibodies, educational materials,...California Davis School of Medicine. This deposit includes frequently cited antibodies for use in brain imaging...disciplines, over time. The resource may also expand to include other affinity reagents, such as nanobodies and...antigen, and performance in various applications are included in the full webpage. Educational Resources...discussed on Addgene’s blog and Antibody Guide, which include information about immunofluorescence, conjugates...over time. Through our incredible community, including your support, we have enabled scientists to easily...

...many roles that actively require these skills, including: Science Reporter Scientific Sales Epidemiologist...Careers where you’ll actively use these skills include: Science Reporter Scientific Illustrator Research...outdoor skills are needed in a variety of roles, including: Livestock Breeder Scientific Illustrator Plant...etc) are highly desirable in a range of roles, including: Science Reporter Technical Writer Statistician...position. Questions you’ll want to find answers to include: Will you be sitting at a desk all day or be on...to the content in your Resume and Cover Letter. Include any relevant job specific ‘buzz words’ to ensure...

...from the company LinkedIn page. This can include news, inclusion, culture and connections. Start your interview... is within a few days after you’ve met. Always include a note about where you met IRL (in real life) and... a connection request to any of these people, including and especially the hiring manager. This is considered.... Follow all the rules of good resume writing. Include examples of concrete accomplishments in both the...the technical and “plus skills” areas. Include a professional picture. It doesn’t have to be a headshot...