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  1. Plasmids 101: Optimizing Plasmid Yields

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    Blog Post
    ... like PCR, cloning, transfection, or long-term storage. But what about those pesky situations where your...maximize cell density and plasmid yield as these factors are generally, but not perfectly, correlated. ...manufacturer. On Addgene plasmid pages, you may see depositor recommendations to grow a plasmid at 30℃ or room...
  2. Genome-wide Screening Using CRISPR

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    Blog Post
    ...population of gRNA-containing lentiviral transfer vectors, each targeting a specific gene within the genome...gRNAs are then cloned into lentiviral transfer vectors to create the CRISPR library. CRISPR libraries ...library that is right for you There are several factors to consider when selecting a CRISPR library for...
  3. Tips for Using BLAST to Verify Plasmids

    Type
    Blog Post
    ...the sequencing results of the plasmids in our repository with known reference sequences, such as full ...full plasmid sequences provided by the laboratories that deposit their plasmids with us or other entries ...entries in NCBI’s numerous databases. As our repository has grown over the years (we now have over 60,000 plasmids...
  4. Which Fluorescent Protein Should I Use?

    Type
    Blog Post
    ...Protein (GFP), a protein from the jellyfish Aequorea Victoria. Since that discovery, the family of FPs just ... used are GECIs - genetically encoded calcium indicators (e.g. GCaMP). Others include: pHluorin & pHTomato...have a filter that will pass blue light to the detector/camera, then BFPs are of no use to you. When using...
  5. Uncovering Ancestral Bias in CRISPR Research

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    Blog Post
    ...Biomedical research has a long and well-documented history of racial bias, which still impacts and influences...Further complicating the issue is the long and ugly history of reporting false essentialist links between biology...actively look for and consider bias as a confounding factor when interpreting findings, particularly when researching...
  6. MXS Chaining

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    Blog Post
    ... construct. This strategy can be used to build vectors with diverse functions, and thus, is used in many...construction of plasmids with many components. The creators of the MXS-chaining method have used the technique...Shetty RP, Endy D, Knight TF. Engineering BioBrick vectors from BioBrick parts. Journal of biological engineering...
  7. Plasmids 101: Colony PCR

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    Blog Post
    ...colonies. The best way to do this is by using your vector with and without an insert to verify that the primers...cultures of your positive clones. If you want to store your clones longer term, just streak the colonies...PCR primers work in the first place: the backbone vector with and without an insert. These controls are ...
  8. Plasmids 101: Protein tags

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    Blog Post
    ...number of chemical and peptide-based proteosome inhibitors, glutathione S-transferase (GST), which can be...purification to get your protein as pure as possible? Vectors that offer different combinations of tags are readily...Protein (GFP) Fluorescent Protein Guide Expression Vectors from the Berkeley QB3 MacroLab Eric thanks his...
  9. Interview: Hodaka Fujii on enChIP, New CRISPR Tools, and More

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    Blog Post
    ...applications? Fujii: We generated a series of retroviral vectors expressing tagged dCas9 and guide RNA. The retroviral...joined Addgene's Advisory Board, as well as the Editorial Board of Scientific Reports and PLoS One. Additionally...especially for young researchers. It was a really nice incubator for me. In Japan, of course, it is optimized for...
  10. Antibodies 101: Immunoprecipitation

    Type
    Blog Post
    ...Western blot. When choosing beads, there are several factors to consider. Agarose beads are sponge-like and ...more stringent buffers may be needed. Since many factors, including salt concentration, ionic strength and...prevent this, include protease and phosphatase inhibitors in the lysis buffer, keep samples on ice, and...
  11. Tips for Writing a Good Cover Letter

    Type
    Blog Post
    ...logistical, like briefly explaining a gap in your job history or sharing relocation plans. They can be skills-based...in the possibility of joining your team as a postdoctoral researcher. Dear Hiring Manager, I am writing...your cover letter, and if possible, send it to a mentor or colleague for a second look and helpful feedback...
  12. Plasmids 101: Codon usage bias

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    Blog Post
    ...- e.g. codon choice may interrupt transcription factor binding sites) As you might imagine, it’s not ...not easy for humans to balance all of these factors on their own. Luckily, many researchers have created...Kimberly A., et al. "Selective charging of tRNA isoacceptors induced by amino‐acid starvation." EMBO reports...
  13. Controlling for Off-target Effects with a New Genome-wide CRISPR Screen Design

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    Blog Post
    ... delivered as a pooled library via a lentiviral vector to cell lines that either stably expressed Cas9... screen did fail to identify some known ricin-regulators, but most of these genes are also essential for...hypersensitivity, or are in an enhancer, transcription factor binding site, or a coding region. Targeting these...
  14. Finding nucleic acids with SHERLOCK and DETECTR

    Type
    Blog Post
    ...time-consuming, and complex to use outside of standard laboratories. In the case of the COVID-19 pandemic, qPCR ...each enzyme. In addition, Csm6, a CRISPR type-III effector nuclease, can be used in conjunction with Cas13...single-component programmable RNA-guided RNA-targeting CRISPR effector. Science 353:aaf5573 . https://doi.org/10.1126...
  15. CRISPR 101: Targeting RNA with Cas13a (C2c2)

    Type
    Blog Post
    ... (LshCas13a) to begin to characterize this new effector (Abudayyeh et al., 2016). In the endogenous CRISPR...fluorophore. The fluorescent signal can be used as an indicator to determine whether the target sequence is present...single-component programmable RNA-guided RNA-targeting CRISPR effector." Science 353(6299) (2016):aaf5573. PubMed PMID...
  16. Fluorescent Proteins 101: Aptamer Fluorophores

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    Blog Post
    ...on FRET techniques. By combining standard donor/acceptor FRET know-how with aptamers, aptabeacons (or aptatags...fluorophore protein. Years later, Samie Jaffrey’s laboratory took advantage of their deep knowledge of the...a higher melting temperature than its Spinach ancestors, and has a lower magnesium dependence. Though ...
  17. Using Video to Share Your Science: We Share Science

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    Blog Post
    ...research scientists, faculty, students, private sector researchers, or pretty much anyone applying the...studies on reducing antibiotic prescriptions for respiratory tract infections. The video abstracts provide...is not hard either. Here are some resources: ImpactStory, a website that helps researchers go beyond citation...
  18. Five Popular Model Organisms

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    Blog Post
    ... al., 2009). By injecting mice with retroviral vectors that harbored different mutations found in leukemia...Gerald Rubin’s lab who created a set of modular vectors that can be used to optimally control gene expression...discovery of RNAi in the worm actually rewarded depositors Andrew Fire and Craig Mello a Nobel Prize in ...
  19. Multicolor Animals: Using Fluorescent Proteins to Understand Single Cell Behavior

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    Blog Post
    ...either by electroporation of plasmids, or using AAV vectors. The Imai lab also developed a version of Tetbow...Livet, Jean, et al. "Transgenic strategies for combinatorial expression of fluorescent proteins in the nervous...Markus Grompe. "Clonal tracing of Sox9+ liver progenitors in mouse oval cell injury." Hepatology 60.1 (...
  20. Plasmids 101: Biotinylation

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    Blog Post
    ... the microflora that reside there. Biotin is a cofactor that in its active form is covalently attached...consists of a biotin molecule attached to a photoreactive arylazide group. Under strong visible light,...single-step purification of tagged transcription factors in mammalian cells and transgenic mice.” PNAS 100.13...
  21. Plasmids 101: Expressing shRNAs for RNA Interference

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    Blog Post
    ...CRISPR, RNAi methods are often quicker and the repositories of validated small RNAs are vast and difficult...processing Following delivery into cells using viral vectors or plasmids, shRNAs hijack the endogenous miRNA...cloning strategies for constructing shRNA expression vectors. BMC Biotechnology, 6(1). https://doi.org/10.1186...
  22. Transferable Skills Guide: Time Management

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    Blog Post
    ...too many activities too close to one another. Monitoring task progress. For this, I always make sure I... lists. Because I know what my goals are (see monitoring task progress), and I know that I’ve blocked ... give it a final read Browse list of previous depositors from Boston University Contact Dr. Gilmore directly...
  23. Site Directed Mutagenesis by PCR

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    Blog Post
    ...introduce or remove restriction sites), in mapping of regulatory elements (to mutate promoters/enhancers in reporter... addition to the “A” site already in the parent vector). Also shown are the “B” restriction sites, which...Browse Our Molecular Biology Protocols Find Empty Vectors for Your Next Cloning Experiment ...
  24. Antibodies 101: The Basics of Western Blotting

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    Blog Post
    ...Aydin S (2015) A short history, principles, and types of ELISA, and our laboratory experience with peptide...knockout line for validation, please view this tutorial from Stuart Orkin's and Daniel Bauer's labs.  ...
  25. Plasmids 101: Dimers and Multimers

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    Blog Post
    ...of multiple circular DNA forms of colicinogenic factor E-1 from Proteus mirabilis. Biochemistry. 1968 ...Guttman A, Thompson L. Vaccine Plasmid Topology Monitoring by Capillary Gel Electrophoresis. Curr Mol Med... JA, Carnes AE, Hodgson CP. Plasmid DNA vaccine vector design: impact on efficacy, safety and upstream...
  26. An “elegans” Approach to Better CRISPR/Cas9 Editing Efficiency

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    Blog Post
    ...modular toolkits build single plasmid targeting vectors that encode the gRNA transcript and the repair ...recognized by the 2020 Nobel Prize in Chemistry), and Victor Ambros and Gary Ruvkun for their seminal work on....114.170423 Schwartz ML, Jorgensen EM (2018) SapTrap vectors for introducing point mutations with unc-119+ selection...
  27. It’s CRISPR Week at Addgene!

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    Blog Post
    ...Now we have over 8,000 CRISPR plasmids in the repository that have been shared over 140,000 times to over... today, we’ll start with a CRISPR overview and history. Tuesday will focus on generating knockouts using...
  28. The Twelve Days of CRISPR

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    Blog Post
    ...first generation of base editors to the eighth generation (and dual base editors, too!).  On the seventh...
  29. SciComm with the Experts at Science in the News Part 1

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    Blog Post
    ...graduate student at Harvard University, former Co-director of Science in the News, and member of the SITN...her PhD from Harvard University, is a former Co-director of Science in the News, and is a member of the...
  30. New Educational Resource: The Addgene Videos Page

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    Blog Post
    ...lab process. Along with our role as a plasmid repository, Addgene aims to be a vital educational resource...the Addgene Blog 5 Science Rap Videos and Their Creator: Tom McFadden SciComm with the Experts at Science...
  31. Podcast: A Malawian Professor's Path to Biotech Research

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    Blog Post
    ...received an award from nonprofit and Addgene collaborator, Seeding Labs, for lab equipment that will help...23:44 : Follow up Kingdom's PhD projects - HIV inhibitors, biofuels, challenges in Malawi, and more 23:...
  32. Call for Guest Bloggers

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    Blog Post
    ...purification Replica plating Anti-CRISPR Retroviral vectors Career development for scientists A new plasmid...do you get out of it? Writing Experience - Our editors will work with you to help hone your blog post ...
  33. CRISPR Cheat Sheet

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    Blog Post
    ... a functional change in the DNA sequence. Base Editors: Modified versions of CRISPR nucleases that make...sequence without completely cutting the DNA. RNA Editors: CRISPR nucleases that cut or modify RNA as opposed...
  34. Alex Chavez on MD PhDs, CRISPR, Gene Drives, & More

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    Blog Post
    ...Pathology at Masachusetts General Hospital and Postdoctoral Fellow in the Church and Collins labs at the... Blog Learn More About Alex's Work with Cas9 Activators Michael Koeris Explains His Path from Grad Student...
  35. Interview with Bart Newland, Addgene’s Newest Board Member

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    Blog Post
    ...introduce the newest member on Addgene’s Board of Directors, Bart Newland. Newland is General Counsel at Atalanta...really just taking off. I found out that patent attorneys with a biology background were in demand. Q: Can...
  36. Addgene’s Blog Gets a New Look!

    Type
    Blog Post
    ...main topics of the blog: Plasmids CRISPR Viral Vectors Fluorescent Proteins Science Careers You can use... us in the comments below. We’ll track down the story, get the inside scoop, and keep you up-to-date. ...
  37. Purple Plants that Report Pollutants

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    Blog Post
    ...specific molecule. By combining this approach with receptors for problematic ligands, crops could report the...available via github, and Dr. Gehan recommends a tutorial on hyperspectral imaging. Step aside, green thumbs...
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