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Showing: 241 - 260 of 960 results

  1. Designing Your Chalk Talk for the Academic Job Interview

    Type
    Blog Post
    ...vision. Practice drawing and writing everything in a form that is easily visible from the back of the room... vision. Organize your vision into a chalk talk format. Practice. Define your research vision A chalk...find, and how you will interpret and use the information. In contrast, a list of experiments or a laundry...Organize your research vision into the chalk talk format Once you have developed your research vision, you...most of your limited speaking time. Beyond the information you seek to convey in your chalk talk, there'...

  2. Tips for a 1st time AAV user (by a Rookie AAV user)

    Type
    Blog Post
    ...are required for proper packaging of AAV, but also form secondary structures that are prone to deletion ...and XmaI restriction sites). Another way is to transform AAV transfer plasmids into bacterial strains, ...found in the culture media, you should consider performing PEG precipitation. Instead of sonicating the ...

  3. Plasmids 101: Codon usage bias

    Type
    Blog Post
    ...choice, you should perform a functional assay to make sure your protein isn’t forming inclusion bodies ...

  4. Negative Can Be Positive: Open AAV Data with Addgene

    Type
    Blog Post
    ...highlights the dearth of AAV vector information provided in openly accessible forms like publications. Why not ... two years testing and reformulating our AAV construct to optimize performance in our system and yet none...literature, and as a result can leave scientists under-informed with respect to how different serotypes, promoters...this one set of experiments didn’t provide any information on other important factors such as what is the...adjacent areas impacted the experiments we wanted to perform. It required a great deal of time and effort to...packaging viral particles (6), can influence vector performance. Open science initiatives, like Addgene’s Data...negative data and instead focus on getting this information out into the public where it can be utilized ...

  5. Antibodies 101: Immunoprecipitation

    Type
    Blog Post
    ...antibody-target protein immune complexes are allowed to form. Following incubation, beads are used to immobilize...-40 or triton-X, to preserve native protein conformations and interactions. For difficult to release proteins... in the lysis buffer, keep samples on ice, and perform the IP at 4oC. When choosing the capture antibody... recognizes the target protein in its native conformation. Frequently, the antibodies used for a standard... are completely removed. Wash steps should be performed even with a pre-cleared sample.  Eluting the target...and negative controls for an IP. When possible, perform the IP in parallel with both a sample known to ...order to identify any non-specific interactions, perform parallel IPs with beads only and an isotype control...

  6. Hot Plasmids - November 2023

    Type
    Blog Post
    ...best PE6 variant for your application. Image reused form Doman and Pandey et al., 2023 under CC-BY 4.0 License.... These variants use different RT enzymes and performed large edits with similar or better efficiency ...potential to reduce negative side effects. This platform offers a promising approach for experimental therapies...we began distribution and has been verified to perform well in standard IHC and western blot protocols...protocols. You can find more specific information on the Wu Lab’s use of Anti-PSD-95 [K28/43R] under the Application...

  7. Plasmids 101: Choosing an Antibiotic Resistance Gene

    Type
    Blog Post
    ... plates. This prevents satellite colonies from forming while effectively selecting plasmid-containing ...cost-effective, timesaving for transformations. Cons: Prone to satellite colony formation on agar plates, less ...in a plasmid. During the propagation and/or transformation process, some bacteria will either never receive... is nifty because the recovery period after transformation (the dreaded 60 minutes shaking at 37 degrees..., interchangeable with Amp, timesaving for transformations. Con: Expensive.   Kanamycin Kanamycin, or...selection gene. However, it does require a slower transformation recovery period than amp or carb (60 minutes...confers mammalian resistance to G418. Con: Slow transformation   Spectinomycin/streptomycin Spectinomycin...

  8. CRISPR 101: Targeting RNA with Cas13a (C2c2)

    Type
    Blog Post
    ... in spacer acquisition following infection, as a form of “bait” to identify new CRISPR-associated proteins...target binding and cleavage through a series of conformational changes in the Cas13a molecule. Like Cas9, ...

  9. Fluorescent Proteins 101: Fluorescent Protein Timers

    Type
    Blog Post
    ...changes in gene expression as new body sections are formed. In synthetic biology, FP timers might be able ...isolate particular organelles and their contents, or perform in situ hybridization experiments. In many cases...like the patterning of nascent tissues and the formation of limbs are correlated with changes in gene expression...and developmental timer of neural fiber bundle formation." Journal of Biological Chemistry 276.32 (2001...

  10. Fluorescent Proteins 101: Aptamer Fluorophores

    Type
    Blog Post
    .... For instance, the fluorophore and quencher may form the base of the stem in an RNA stem-loop structure...and detection - since they frequently undergo conformational changes after target binding, aptamers are ...fluorescence-activated cell sorting (FACS) of E. coli transformed with aptamer libraries, the Jaffrey group selected...

  11. CRISPR Between the Genes: How to Experiment with Enhancers and Epigenomics

    Type
    Blog Post
    ...compaction—on the order of 700 terabytes per gram—and form the X-shaped chromosomes we all know and love. To...enhancers and genome math come in. They help us to transform raw genomic coordinates into meaningful experiments...tri_methylation In the next section we’ll apply the above formula to a real-world experiment. We’ll start with ChIP-seq...

  12. Minigenomes - a Safe Way to Study Dangerous Viruses Like the Ebola Virus

    Type
    Blog Post
    ... associates with the viral polymerase, which is formed by a complex of two proteins: the large (L) catalytic...measured to quantify (indirectly) transcription performed by the viral polymerase. Alternatively, the mRNA...independently. In addition, experiments can be performed in the absence of some viral proteins (compared... minigenome plasmid per well (in a 6 well plate format). However, since plasmid quality and transfection...

  13. Avoiding the Dark Side of Fluorescent Protein Fusions with mOX FPs

    Type
    Blog Post
    ...and others have reported that FP cysteine residues form inappropriate disulfide bonds in the secretory pathway...modifications including glycosylation, disulfide bond formation, and proteolytic cleavage. FPs localized to organelles...fusion protein localization. For example, the formation of inappropriate inter-chain disulfide bonds between...because of the correct folding and fluorophore formation by most, if not all, of the FP fusions. Our moxFPs...

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

    Type
    Blog Post
    ...Barcoded tubes come in many forms. The 2D barcode can store a lot of information in a small space and is ... treated to easily take in plasmids (transformation). Transformed bacteria are then grown up on antibiotics...received is immediately transformed and stored. Some samples need to be transformed with a micropulser using... (98.2%) transform the first time and can be stored right away. Every week, the lab performs an average...average of 140 transformations. The lab typically transforms newly deposited samples from Monday through Wednesday...delivered by a mail courier and then the journey of transformation and storage begins. Some samples are submitted...barcoded tubes that are scanned into our Laboratory Information Management System (LIMS), a database we use to...

  15. Plasmids 101: Protein tags

    Type
    Blog Post
    ...can also protect against proteolysis. That’s one form of instability. Prokaryotes can also have a hard...Thioredoxin A (TrxA) that improve disulfide bond formation in order to help keep your protein soluble.  ... better assess their function. More detailed information and some protocols can be found in the references...

  16. Negotiating Work and Life: How to Find the Joy

    Type
    Blog Post
    ... local family to help, create “fake” family by forming a social circle of people in the same situation... and/or your spouse won’t be at every school performance, chaperone every field trip or cheer at every...

  17. Arf GTPase Family

    Type
    Collection
    ... RAS superfamily, though now have been argued to form their own superfamily (Klinger, et al., 2016). Though...added to this list? Fill out our Suggest a Plasmid form or e-mail [email protected] to help us improve this... table lists genes in the ARF GTPase family, information on each gene, and examples of plasmid types available...table lists genes in the ARF GAP and GEF family, information on each gene, and examples of plasmid types available...
Showing: 241 - 260 of 960 results