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  1. Light Sheet Fluorescence Microscopy

    Type
    Blog Post
    Published
    July 16, 2015, 2:30 p.m.
    ...The underlying method was developed in 1902 by Richard Zsigmondy and Henry Siedentopf to enhance the microscopic...using cameras with electron multiplying couple-charged device (EMCCD) or complementary metal-oxide semiconductor...

  2. Isolating Genomic Regions of Interest with the CRISPR System

    Type
    Blog Post
    Published
    May 30, 2019, 12:44 p.m.
    ...identification of DNA binding proteins, this method can characterize nucleic acid-nucleic acid interactions at a ...2017). This upregulates expression of IGF2, a characteristic of many human malignancies. Additionally, the...

  3. Simplify Cloning with in vivo Assembly

    Type
    Blog Post
    Published
    Oct. 18, 2018, 12:37 p.m.
    ...and likely functions as a bacterial DNA repair mechanism. The pathway is ubiquitous, with successful recombination...placing homologous ends on DNA using polymerase chain reaction." Biotechniques 10.1 (1991): 62-66. PubMed...

  4. AAVs in Retinal Gene Therapy

    Type
    Blog Post
    Published
    July 31, 2018, 12:55 p.m.
    ...occurring animal model, the RPE65−/− dog, was characterized and found to suffer from early and severe visual.... PubMed PMID: 29983335. 9. Day TP1, Byrne LC, Schaffer DV, Flannery JG, 2014. Advances in AAV vector ...

  7. DNA Purification Without a Kit

    Type
    Blog Post
    Published
    Feb. 11, 2020, 2:15 p.m.
    ...but the addition of a high concentration of a chaotropic salt (guanidine hydrochloride for the plasmid... glass milk, gel slices are first placed in a chaotropic salt sodium iodine (NaI) solution. NaI has two...

  8. A Guide to Starting Your Own Journal Club

    Type
    Blog Post
    Published
    Feb. 6, 2020, 3 p.m.
    ...journal club should aim to have many of the same characteristics as a successful meeting: a clear purpose and...success” of the journal club. This can be a challengingly vague metric but there are a few things you ...

  9. Viral Vectors 101: The Retroviral Lifecycle

    Type
    Blog Post
    Published
    July 27, 2023, 1:15 p.m.
    ...genomic sequence from one another) — by various mechanisms, only one of the two strands’ DNA counterparts...γ-Retrovirus Guide References Cullen, B. R. (1992). Mechanism of action of regulatory proteins encoded by complex...

  10. Great Results Start with Great Standard Curves

    Type
    Blog Post
    Published
    Aug. 15, 2024, 1:15 p.m.
    ...final readout. When preparing a standard curve, change pipette tips and mix thoroughly by inversion or...dilution series, use consistent dilution factors, change pipette tips regularly, and mix samples very well...

  11. Uncovering Ancestral Bias in CRISPR Research

    Type
    Blog Post
    Published
    Nov. 29, 2022, 2:15 p.m.
    ...Cancer Data Science team at Broad, they were able to change their analysis to use a corrected genome map. Now...): 250-252. Carlson J, Henn BM, Al-Hindi DR, Ramachandran S. Counter the weaponization of genetics research...

  12. CRISPR 101: Ribonucleoprotein (RNP) Delivery

    Type
    Blog Post
    Published
    Sept. 6, 2018, 12:02 p.m.
    ...generation of the RNP complex.  While one option is to purchase the Cas9 protein and a gRNA from a commercial ...performed rapidly and, in many cases, with reduced chance for off-target effects. As the CRISPR field continues...

  13. Better Dyeing Through Chemistry & Small Molecule Fluorophores

    Type
    Blog Post
    Published
    Sept. 8, 2016, 2:30 p.m.
    ...college student searching couch cushions for spare change, desperate to extract a few more photons from a...Fluorescent Protein Fusions Request Plasmids from the Michael Davidson Collection Find Fluorescent Protein Kits...

  15. Tips from the Repository Trenches: Using Barcodes to Track Samples

    Type
    Blog Post
    Published
    Oct. 6, 2015, 2:30 p.m.
    ...transformed with a micropulser using an electric charge to help the DNA enter the cells. Some samples require... to be read one at a time. 1D barcodes can be purchased from many companies such as Corning. Tubes can...

  16. The Importance of a Fun Workplace: Company Culture at Addgene

    Type
    Blog Post
    Published
    Sept. 3, 2015, 2:30 p.m.
    ... included a simple corn hole set and a cooler. Chances are one of your colleagues has a frisbee at least...where to be and when. This pre-planning will avoid chaos and delays in getting the fun started. I’m aware...

  17. Your Lentiviral Plasmid FAQs Answered

    Type
    Blog Post
    Published
    April 23, 2014, 1:08 p.m.
    ...lentiviral and retroviral packaging vectors are not interchangeable. While these differences may seem subtle they...the generation used to package the virus does not change the generation of the transfer vector. Q5: What...

  18. Plasmids 101: Gateway Cloning

    Type
    Blog Post
    Published
    Jan. 12, 2017, 3:30 p.m.
    ...destination vectors prior to recombination, and it is exchanged with the gene of interest during the BP or LR ...Multiple Hosts. Curr Protoc Protein Sci. 2003 Feb;Chapter 5:Unit 5.17. PubMed PMID:18429245. 3. Ptashne, ...

  19. Mesothelioma - Causes, Symptoms, and Treatment

    Type
    Blog Post
    Published
    Dec. 13, 2016, 3:30 p.m.
    ...Although different types of cancer have different challenges and obstacles to overcome, mesothelioma sufferers...of immunotherapy is the use of drugs (or other mechanisms) that work through body’s immune system to fight...

  20. Plasmids 101: Plasmid Incompatibility

    Type
    Blog Post
    Published
    March 31, 2020, 1:15 p.m.
    ...during cell division. They don’t simply rely on chance (or random diffusion) to make this happen.  There...centromere-based incompatibilities: Insights into the mechanism of incompatibility mediated by low-copy number...

  21. Viral Vectors 101: Pseudotyping

    Type
    Blog Post
    Published
    April 20, 2021, 2:30 p.m.
    ...have different levels of cytotoxicity and the mechanisms behind the cytotoxicity can vary. For example...differentially inactivated by nonspecific complement mechanisms in serum. Sometimes this is due to the cells ...

  22. CRISPR 101: Which Cas9 Do I Choose for My CRISPR Experiment?

    Type
    Blog Post
    Published
    Oct. 13, 2022, 1:45 p.m.
    ... been converted to an alanine (D10A or H840A), changing its double-strand cut to a single-stranded nick...Walton et al., 2020). Further, this group also characterized a second Cas9 variant, SpRY, which cuts at NRN...

  23. Rett Syndrome: A History of Research and Therapeutic Outlooks

    Type
    Blog Post
    Published
    Nov. 19, 2020, 2:15 p.m.
    ... Rett Syndrome, respectively. One of the main challenges facing Rett researchers is how to deliver the... by Gail Mandel. There are numerous technical challenges to overcome for both of these approaches, such...

  24. Plasmids 101: E. coli Strains for Protein Expression

    Type
    Blog Post
    Published
    Feb. 10, 2015, 3:06 p.m.
    ...suited for recombinant protein expression. Many challenges can arise when over-expressing a foreign protein...minutes before adding the inducer. Growth Media: Changing media is tricky, because there can be a trade-...

  25. SARS-CoV-2/COVID-19 Detection Methods Based on CRISPR/Cas

    Type
    Blog Post
    Published
    May 5, 2020, 1:15 p.m.
    ... This test can be performed in two steps for challenging samples as in Two-Step-SHERLOCK or in a fast,...the immobilised target sequences. Upon binding, changes in fluorescence can be visualised. Another approach...

  26. Antibodies 101: Introduction to Gating in Flow Cytometry

    Type
    Blog Post
    Published
    April 4, 2024, 1:15 p.m.
    ...ll notice that while the height of your events changes, the separation pattern of positive vs. negative...live population from my live-dead staining. (B) Changing the y-axis from SSC to FSC can alter the positioning...

  27. CRISPR 101: Validating Your Genome Edit

    Type
    Blog Post
    Published
    Nov. 3, 2022, 12:15 p.m.
    ...enzyme screening is sensitive to single base pair changes, but the edit must occur at an existing restriction...simultaneously validate edits and look for off-target changes. This method requires a set of control cells to...

  29. Site Directed Mutagenesis by PCR

    Type
    Blog Post
    Published
    Aug. 2, 2016, 2:30 p.m.
    ...role of renaturation temperature in the polymerase chain reaction." Analytical Biochemistry 209.2 (1993):...Xia, Yongzhen, et al. "New insights into the QuikChangeTM process guide the use of Phusion DNA polymerase...

  30. Targeting HIV-1 with CRISPR: Shock and Kill or Cut it Out?

    Type
    Blog Post
    Published
    May 10, 2016, 2:30 p.m.
    ... a well-established viral reservoir. A second challenge for CRISPR HIV-1 therapies is sequence specificity.... Ramanan V, Shlomai A, Cox DB, Schwartz RE, Michailidis E, Bhatta A, Scott DA, Zhang F, Rice CM, Bhatia...

  32. Plasmids 101: Common Lab E. coli Strains

    Type
    Blog Post
    Published
    Nov. 7, 2014, 2:56 p.m.
    ...below outlines a few of the more common genetic changes found in E. coli strains. Table 1: Common gene ...are methylation sensitive. dnaJ Mutation in a chaperonin gene Increases the stability of certain expressed...

  33. How to Design Your gRNA for CRISPR Genome Editing

    Type
    Blog Post
    Published
    Sept. 24, 2020, 1:15 p.m.
    ...the C’ terminus of the protein, to maximize the chances of creating a non-functional allele. For a 1 kilobase...the so-called base editor Cas9, which makes DNA changes in the absence of dsDNA breaks (Rees et al., 2018...

  34. 9 tips for a successful postdoctoral experience

    Type
    Blog Post
    Published
    Oct. 9, 2018, 12:21 p.m.
    ...Today's postdoctoral fellows (PFs) face a number of challenges ranging from long periods of training to limited...is equally important to have a plan to get to mechanism. Do the necessary tools exist to tackle the problem...

  35. Transferable Skills Guide: Teamwork

    Type
    Blog Post
    Published
    Nov. 28, 2017, 2 p.m.
    ...Watson, Crick, & Franklin, Meselson-Stahl, Hershey-Chase...Who was I to dispute history? And boy did I make...process that kits require; Nicole, the web master in charge of all our externally facing kit pages. No single...

  36. Quick Guide to All Things Lentivirus

    Type
    Blog Post
    Published
    March 21, 2017, 2:30 p.m.
    ... using proper technique, there’s always a small chance that a researcher could infect him or herself with...you put in your culture. As viral production can change from one batch to another. For experimental consistency...

  37. Tips for a 1st Time CRISPR User (by a 1st Time CRISPR User)

    Type
    Blog Post
    Published
    March 7, 2017, 3:30 p.m.
    ... genomic DNA to amplify the gRNA targets was challenging because you’re limited by the sequence of the...Vaimberg EW, Donovan KF, Smith I, Tothova Z, Wilen C, Orchard R, Virgin HW, Listgarten J, Root DE. Nat Biotechnol...

  38. A Guide to Getting Started in Undergrad Research

    Type
    Blog Post
    Published
    June 8, 2021, 1:15 p.m.
    ...were inspired by the imminent threat of climate change to study plant and marine biology, environmental...and while doing scientific research. It’s okay to change your mind about what exactly you want to do—I started...

  39. CRISPR 101: Multiplex Expression of gRNAs

    Type
    Blog Post
    Published
    Sept. 10, 2020, 11:45 a.m.
    ... editing or labeling efficiency doesn’t usually change when you add multiple gRNAs on one plasmid. Sound...contains all of the desired gRNAs. This increases the chance that all the edits you want to make will happen...

  40. How to Deposit Your Plasmids with Addgene

    Type
    Blog Post
    Published
    Feb. 27, 2024, 2:15 p.m.
    ...indicate the amino acid change as well as any functional consequences of that change. For example, "Mutated...
Showing: 601 - 640 of 793 results