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Showing: 741 - 760 of 960 results
  1. CRISPR 101: Which Cas9 Do I Choose for My CRISPR Experiment?

    Type
    Blog Post
    ...Motif (PAM), the sequence NGG.  If these conditions apply to your experimental target, SpCas9 and its enhanced...specificity over a single SpCas9. The drawback of this approach is that two sgRNAs targeting opposite strands ...
  2. CRISPR Activation: A Practical Guide

    Type
    Blog Post
    ...suffer from an inability to easily deliver the appropriate isoform(s) of a given gene, as, many times, the...engineering parameters via a library-on-library approach. Nat Methods 12:823–826 . https://doi.org/10.1038...
  3. Plasmids 101: Control Plasmids

    Type
    Blog Post
    ...RNAi or CRISPR, non-targeting controls may be appropriate as they allow you to assess the specificity of...us now revisit our experiment. In Figure 1, it appeared as though the shRNA did not knock down expression...
  4. Plasmids 101: E. coli Strains for Protein Expression

    Type
    Blog Post
    ...plasmid-- make sure to maintain this by growing on appropriate media. Note: Purifying your expression plasmid...to make sure your cells have first reached an appropriate density. Cells in the exponential growth phase...
  5. Using a Light Microscope Protocol

    Type
    Protocol
    ...see your image through both sides. Once you are happy with the lighting, use the coarse focus knob to ...make minor adjustments to the focus. After you are happy with the positioning and focus of your image on ...microscope after use and store the microscope appropriately....
  6. Plasmids 101: Expressing shRNAs for RNA Interference

    Type
    Blog Post
    ...world, there is a myriad of tools this could be applied to. When it comes to altering gene expression, ...is an umbrella term for a number of different applications that involve different types of small RNA molecules...
  7. Antibodies 101: Introduction to Gating in Flow Cytometry

    Type
    Blog Post
    ...adjust this setting as needed. To figure out the appropriate voltages to observe your cells on a flow plot...voltage describes an electric potential that can be applied to the photomultiplier inside the cytometer to ...
  8. Deep Mutational Scanning with One Pot Saturation Mutagenesis

    Type
    Blog Post
    ...that expresses all desired variants of a protein. Applying a selective pressure winnows the pool down to ...technique described in Wrenbeck et al is a PCR-based approach for generating a customizable comprehensive mutagenesis...
  9. Fluorescent Proteins 101: When GFP lets you down

    Type
    Blog Post
    ... proteins.Therefore, GFP is not suitable for applications that require anaerobic conditions. In experiments...protein module that can report on location. This approach enables the study of proteins in cells, but not...
  10. Transferable Skills Guide: Time Management

    Type
    Blog Post
    ... cell splitting issue at all if I’d planned appropriately. Back in my undergrad days, I had general goals... do I write things down? It’s more viscerally appealing for me to physically cross things out (it just...
  11. Five Popular Model Organisms, Part 2

    Type
    Blog Post
    ...Escherichia coli as a Model Organism and Its Application in Biotechnology. In Escherichia coli - Recent...Physiology, Pathogenesis and Biotechnological Applications. InTech. https://doi.org/10.5772/67306 Koornneef...
  12. How to Negotiate a Successful Lab Start Up Budget

    Type
    Blog Post
    ...discounts, collaborations, access to internal award applications, etc. If this is the institute that can help...budgets and invite you to leave a comment below Happy Sciencing, Damien   Damien Wilpitz is a laboratory...
  13. Handling Plasmids from Addgene - Purifying Plasmid DNA

    Type
    Protocol
    ...overnight culture of bacteria . Pro-Tip Refer to appropriate DNA prep protocol for volume of bacteria to grow...the plasmid DNA. Either spin to pellet the DNA or apply the solution to a column that will bind the now ...Aqueous DNA phase Middle phase - A white layer may appear, consisting of precipitated protein particles Bottom...
  14. Plasmids 101: Dimers and Multimers

    Type
    Blog Post
    ...something odd in our uncut DNA control: a band or two appear on an agarose gel at notably higher molecular weights... only in the uncut DNA; the rest of the digest appears normal and produces the expected fragments for ...
  15. Antibodies 101: Antibody Engineering and Directed Evolution

    Type
    Blog Post
    ...extensively characterized and validated, with the application data from IPI and other labs easily viewed in...2021). Directed Evolution: Methodologies and applications. Chemical Reviews, 121(20), 12384–12444. https...
  16. Cancer and the Immune System: Deciphering the Relationship

    Type
    Blog Post
    ...immunotherapeutic agents. Currently the FDA has 12 approved monoclonal antibodies targeting the PD-L1 pathway...Dendreon Corporation is currently the only FDA-approved DC therapy for prostate cancer and is aimed at...
  17. Lentivirus ddPCR Titration

    Type
    Protocol
    ...complete: Prepare 7 microcentrifuge tubes with the appropriate amount of media volume listed in the chart below...tube. Add 4 µL of the 25 ng/µL samples to the appropriate PCR tubes. Pipette back and forth 10 times to...the average infectious titer obtained from the appropriate dilutions to calculate the final infectious titer...
  18. CRISPR/Cas9 FAQs Answered!

    Type
    Blog Post
    ...with the newest CRISPR technologies and their applications can be exhausting. A quick search for "CRISPR...Laganière, J., Cheng, A. W., Hockemeyer, D., Gao, Q., Alagappan, R., Khurana, V., Golbe, L. I., Myers, R. H., ...
  19. Protocol - How to Perform Sequence Analysis

    Type
    Protocol
    ...within the entire construct. Is the primer an appropriate distance from the target? The primer should be... should I do? Check your trace file first; the apparent mismatch/mutation may be the result of a mis-called...
  20. Water Bath Protocol

    Type
    Protocol
    ...in place. Turn the water bath on and set the appropriate temperature as described in your protocol. How...in step 3. You will also need to maintain the appropriate water level in the water bath so before use, ...
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