...without scrolling up and down the page.  Figure 3: The Education & Tools menu Further down What happens if ...and easily navigate to educational resources, tools, collections in our catalog, and the Addgene Help ...search bar. And if you’re in the mood to browse our catalog, you can use the links under the search box to ... mission, links to our deposit, request, and educational resources page, and previews for our three most...

...  Fig. 3: Restriction enzyme screening for resistivity to digestion, indicating targeted edits... originally written by Melina Fan and updated Nov 3, 2022 by Susanna Stroik. You’ve designed your gRNA...how many copies you are looking for. Secondly, categorize your edit – is it any frameshift mutation? A ...a manageable number of the higher probability locations. If you’re worried about off-target effects, you...Tao Chen, Jos Jonkers, Bas van Steensel. Easy quantification of template-directed CRISPR-Cas9 editing. (...and variant-aware in silico off-target site identification for CRISPR genome editing. (2019) Bioinformatics...

...back to the drawing board.  Cycle 3: A Return to HQ Figure 3: Even Blugene is practicing social...Creating great educational science videos is tough. Is the content accurate? Are we conveying complex... had set up involved multiple people in tight locations. Conference rooms for on-screen presenters! Labs...wasn’t going to work for the majority of our educational videos. In addition, it’s really asking a lot...receiver, to beam the camera’s image to my remote location, live. After doing a bit of looking, I found an...

...up the experiment. The first way, it will take you 3 days to complete the entire experiment, but you’ll...be quite difficult. In my experience, planning has 3 components: A mechanism for blocking off time for... the planning section below, it’s important to dedicate specific amounts of time to specific tasks beforehand...instance, if I don’t have draft blog post ready for publication for tomorrow, I’d better upload that post from...handle doing a particularly annoying task and then dedicating that much time to it each day. Once you get over...Student Community Learn about Careers in Science Communication Resources at Addgene.org Check out Careers...

... of how much you can expect to use in the future. 3. Identify the skills needed to use these tools Outline...then it’s worth working within their limitations. 3. Always provide value to others This is a non-profit.... After identifying all the major equipment, categorize all of your minor tools, like pipets, or mini-centrifuges...titles in the regions where the host institute is located. For example, salaries in Kansas versus salaries... most start-up P.I.’s. It doesn’t have to be complicated, or difficult to create this list. Start BEFORE...discounts, collaborations, access to internal award applications, etc. If this is the institute that can help...

... back to a pluripotent stage (iPSC formation) [2, 3]. The iPSCs then proliferate and redifferentiate to..., 2007. 131(5): p. 861-72. PubMed PMID: 18035408. 3. Takahashi, K. and S. Yamanaka, Induction of pluripotent...converting one cell into another. Cell Stem Cell, 2008. 3(4): p. 382-8. PubMed PMID: 18940730. Additional Resources...joined Cell Press to pursue a career in science communication and publishing after completing her postdoctoral...PMC4096605. 30. Lamba, D.A., et al., Generation, purification and transplantation of photoreceptors derived...

...research, training, startups, and real applications of SynBio. Chapter 3: Synthetic biology as the engineering...synthetic genome project and constructing ever more complicated biological devices. Chapter 6: - Making research...Addgene Blog Learn about the Synthetic Biology Education Nonprofit BioBuilder Plasmids 101: Golden Gate...

...PMC4320685. 3. Fu, Yanfang, et al. "Improving CRISPR-Cas nuclease specificity using truncated guide RNAs...negative controls for genome-wide CRISPR screens; and 3) safe-targeting guides, which target genomic sites... potential control, and provide evidence that truncated gRNAs have improved specificity with only a small...toxicity in high-throughput screens." Nature communications 8 (2017): 15178. Pubmed PMID: 28474669. Pubmed...

... sensing!   Figure 3: Components of the types of voltage indicators reviewed.   References...Voltage indicators are voltmeters for cells! In this blog we will review how these indicators work, their...fluorescence or voltage indicators.   Viral Vectors and Voltage Indicators As previously mentioned...Enhanced genetically encoded voltage indicators advance their applications in neuroscience. Curr Opin Biomed...Cellular communication and function depend on potential energy. Cellular activities like action potentials...their general applications, and why viruses are essential to their use as biological tools. Why Use Voltage...Voltage Indicators? Electrical signals are integral to many facets of biology. For example, many heart...

...quantitative real-time PCR. J Microbiol Methods. 2006 Jun;65(3):476-87. doi: 10.1016/j.mimet.2005.09.007. Epub 2005...run in triplicate.  The nice thing about qPCR is that since it measures relative quantification - that... not need to be run in triplicates. ddPCR is an extremely accurate application, even in areas of the genome...Digital PCR for AAV QuantificationPolymerase Chain Reaction Overview and Applications Resources on Addgene.org...blog post, we're going to focus on PCR-based applications (qPCR and ddPCR.) qPCR Quantitative/real-time...range of RQ values as noted above.   In both applications, results can be affected by DNA quality and ...absolute quantitative data, instead of relative quantification.  Even though no one option is perfect, with...

...228, no. 2 (2012): 267-75. PubMed PMID: 22767332.  3. Kim, Dohoon, Chun-Hyung Kim, Jung-Il Moon, Young-...PubMed PMID: 19337237. PubMed Central PMCID: 2677165. 3. Jia, Fangjun, Kitchener D. Wilson, Ning Sun, Deepak... deriving human iPS cells." Nature Methods 7, no. 3 (2010): 197-99. PubMed PMID: 20139967. PubMed Central...serially transfected using a cationic vehicle. The following synthetic modifications are used to protect the... to verify there’s no genomic integration Replicating EBNA1 episome Non-integrating Medium Medium ...creates heterogeneous iPSC cell lines, which can complicate comparisons made between lines. Incomplete silencing...Retroviruses are not considered safe for clinical applications.     Lentiviral: Lentiviral vectors infect dividing...

...Originally published Mar 3, 2016 and last updated Apr 13, 2021 by Will Arnold. Although CRISPR systems...a subsequent curing protocol that requires only 2-3 hours incubation. E. coli and T. citrea scarless editing...the QCascade DNA-targeting module with the crRNA, (3) three transposase proteins, and (4) the donor DNA...for successful modifications, efficiency can be low, especially for larger modifications. What’s the solution...several potential CRISPR applications. First, CRISPR can be used to target locations in the genomes of pathogenic...tools available for bacteria and some of the applications for which they’ve been used. The beginnings ... of Enterobacteriaceae, without the need for modification. This finding suggests that the system may be...

...individual, sharp and evenly spaced peaks …like these…. 3. Expect to get 500-700 bases of clean reliable DNA...of interest. 5. Use a silica spin column for purification of the samples you send for DNA sequencing If...requires you to perform your own Big Dye PCR amplification reaction (as opposed to using the all inclusive...

...trying to raise the alpha isoform of GSK-3 to the same (deserved) stature as the beta isoform. Follow ...where readers may locate data supporting the research reported in PLOS publications. The policy was not... is founded on the principle that scientific publications must contain enough information (or links to...probable to some extent - but that is the price of publication. Who is to say that the initial data-generating...logically annotated and arranged, as in an actual publication and not require interaction with the data authors...likely, at least in part, due to our inadequate cataloguing of how results were obtained and this alone is... Nature Publishing Group recently launched a publication called Scientific Data, which aims to act as ...

...with an isotype-matched control.      Figure 3: A preimmune serum control addresses situations where....” British Journal of Pharmacology, vol. 175, no. 3, Wiley, 19 Jan. 2018, pp. 407–411. Crossref, doi:10.1111...results.  Replicates Replicates are another critical control in biological experiments. Replicates can be...of biological replicates such as multiple mice in each group and C, technical replicates or repeated measurements...secondary antibodies.   Application-specific controls Some antibody-based applications have very specific ...then moving on to controls for antibody-related applications. After reading, you will have a better understanding...blot and look for the molecular weight shifts indicative of tau phosphorylation. By including the positive...

... fast indicator evolved for two-photon microscopy. Cell. 2022 Aug 16. pii: S0092-8674(22)00916-3. doi:...available as viral vectors (here and here).  Figure 3: JEDI-2P was identified from a high-throughput screen...sensors One of the latest additions to Addgene’s catalog is a new class of sensors for neuropeptides and...doi: 10.1038/s41592-022-01597-x. New glutamate indicator iGluSnFR3 Improving on the glutamate sensor iGluSnFR...They generated variants optimized for different applications (v82 has higher affinity for glutamate but slower...PDGFR.codonopt    Aggarwal A, et al. Glutamate indicators with improved activation kinetics and localization...have developed a new genetically-encoded voltage indicator (GEVI) called JEDI-2P (Liu et al. 2022) optimized...

...are 24 variants of the cas14 gene that cluster into 3 subgroups (cas14a-c). All these variants share the...of dsDNA and RNA in trans. Fluorescent signal indicates the presence of this viral sequence in the assay...

...cloning method to human proteins known as Pumilio/fem-3 mRNA binding factors (PUF). In a single step, researchers...says the new tool might even find therapeutic application in addition to their use in basic biology, for...

...to allow for antibodies to enter the cell (Figure 3). A useful side effect of the fixing/permeabilization...cytosolic procedure in that case.     Figure 3: For intracellular labeling cells need to be fixed...labeling can be subdivided into two categories, depending on the location of the targeted markers. Some commercially...proteins, such as receptors, or post-translational modifications on those proteins, like glycosylation patterns...Depending on the research question and downstream application, one or several of those techniques can be used...your flow analysis. However, certain downstream applications, like an in vivo adoptive transfer of those ...labeling Oftentimes, a marker of interest is not located on the cell membrane but inside the cell. Due to...

... 5’ and 3’ LTRs and expressed them along with Cas9 in the T-cell line 2D10. PCR amplification and Sanger...strategies: potential tools for eradicating HIV-1/AIDS. J Neurovirol. 2015 Jun;21(3):310-21. PubMed PMID: 25716921...pre-treatment levels due to the viral reservoirs located in latently infected cells. Although ART-treated...toxic proteins, CRISPR/Cas9 SAM caused apoptosis, indicating a true “shock and kill” response. Cutting HIV...create mutations in genes essential to HIV-1 replication. However, this approach could permit some viral... HIV-1 could escape from CRISPR/Cas9-induced modification targeting either the LTRs or essential genes...genes. Many of these escape mutations were located near the Cas9 cut site, leading Wang et al. to conclude...