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Showing: 721 - 740 of 949 results
  1. Hot Plasmids - August 2020

    Type
    Blog Post
    ... have created a yeast-based directed evolution approach with some notable improvements. Directed evolution...specific guide RNA (gRNA) sequences. But these approaches rely on continual expression of both dCas9 and...
  2. 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 ...
  3. Viral Production at Addgene

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    Blog Post
    ...particles can be identified after negative staining and appear darker than full vector particles.     ...replaced with a new vial of early passage cells. Approximately two weeks post-thaw, cell culture supernatant...
  4. How to Be an Excellent Trainee

    Type
    Blog Post
    ...easiest to train individuals who are polite, appreciative, and clear about their needs. Remember, your...trainees one day when you are the senior personnel! Happy training! Resources Resources on the Addgene blog...
  5. QC Sequencing Technologies at Addgene

    Type
    Blog Post
    ...bioinformatic analysis. At Addgene, we use a mix-and-match approach for sequencing methods and choose the best method...Nanopore sequencing technology, bioinformatics and applications. Nature Biotechnology, 39(11), 1348–1365. https...
  6. Antibiotic Resistance: An Old Solution but a New Problem

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    Blog Post
    ...from random mutations in the genetic sequence that happen to be beneficial, and persist as a result. Why ...are susceptible to the drug, while the few that happen to possess resistance genes survive and multiply...
  7. CRISPR Activation: A Practical Guide

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    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...
  8. Plasmids 101: Control Plasmids

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

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    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...
  10. 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....
  11. Handling Plasmids from Addgene - Purifying Plasmid DNA

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    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...
  12. 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...
  13. 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...
  14. 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 ...
  15. 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...
  16. Plasmids 101: Expressing shRNAs for RNA Interference

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    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...
Showing: 721 - 740 of 949 results