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We narrowed to 949 results for: ache

Showing: 521 - 540 of 949 results

  1. Hot Plasmids - February 2022

    Type
    Blog Post
    ...020-0600-6. The Penn State Protein Ladder  by: Rachel Leeson This simple, inexpensive plasmid-based ladder... which also provides a useful yet low-pressure teaching opportunity for lab members or students new to...for mammalian genome regulation and editing by: Rachel Leeson The large size of CRISPR-Cas effectors and...

  3. Q&A with Karen Katz and Carl Paratore, the Newest Members of the Addgene Board of Directors

    Type
    Blog Post
    ...with Addgene? Katz: Joanne and I first got to know each other through the Healthcare Businesswomen’s Association...Paratore: Kristen Darby, who’s also a board member, reached out and asked if I would be interested in joining...responsible for bringing a systematic and disciplined approach to evaluating and improving the effectiveness ...

  4. Cas13d: Small RNA-targeting CRISPR enzymes for transcriptome engineering

    Type
    Blog Post
    ...CRISPR array yielded >90% knockdown by CasRx for each of four targets, including two mRNAs and two nuclear...which efficiency varies widely across guide RNAs, each guide tested with CasRx yielded >80% knockdown. ... exon skipping. Previous CRISPR exon-skipping approaches used two guide RNAs to remove a given exon from...

  5. Viral Vectors 101: Viral Vector Elements

    Type
    Blog Post
    ...First, it needs to be virally enveloped which is achieved by expression of the vesicular somatitis virus...within the viral particle. The ITRs base pair with each other to allow DNA synthesis of the complementary...and gamma-retroviruses have a lot in common with each other. Below we will outline the common features...

  6. Multiplex Genome Editing with CRISPR-Cpf1

    Type
    Blog Post
    ...methods relied solely on Cas9. Overall, these approaches have two main drawbacks: 1) Most rely on transfection... One vector expresses Cas9 and up to 7 gRNAs Each gRNA requires its own promoter Sakuma et al   ...One vector expresses Cas9, eGFP, and up to 4 gRNAs Each gRNA requires its own promoter   Kabadi et al ...

  7. Clear and Accessible Writing

    Type
    Blog Post
    ...a lot of jargon and have a technical, detailed approach. Other times, it is more helpful to use plain ...text for their practice image. We know; we asked each group to share with the rest of the workshop.  For...muddled due to the passage of supernatant through each layer. The right panel illustrates a gradient tube...

  8. Plasmids 101: Screens vs. Selections

    Type
    Blog Post
    ... are here to discuss the ins and outs of both approaches. Screens and selections The common goals The...details - and that’s where the differences in these approaches can be found. As an example, say you are considering...and consider these questions to save yourself a headache later on. Enhancer and suppressor screens These...

  9. CRISPR 101: Non-Homologous End Joining

    Type
    Blog Post
    ..., repressing RecA and overexpressing the NHEJ machinery improved NHEJ accuracy in M. smegmatis (Yan et...also be engaged by variants of the canonical Cas9 approach. A pair of CRISPR guides that flank regions of... break; when two such breaks are introduced near each other, in opposite strands. This “double nickase...

  10. Plasmids 101: Yeast Vectors

    Type
    Blog Post
    ...widely utilized across many different cell types and each one requires different elements for vector propagation...control not only the number of plasmids found in each cell, but also whether the plasmid gets integrated...non-physiological levels. Scientists have tried varied approaches to combat these issues. One method to reduce ...

  11. Plasmid Cloning by Restriction Enzyme Digest (with Protocols)

    Type
    Protocol
    ...it is also a good idea to run an uncut sample of each plasmid to help with troubleshooting if your digests...approximately 1:3. Since the number of base pairs for each varies, it is difficult to calculate this based ...alone. One method is to conduct 2 ligations for each plasmid you are trying to create, with varying ratios...

  12. Antibodies 101: Selecting the Right Antibody

    Type
    Blog Post
    ...you can easily distinguish each different type of antibody and therefore each target protein. This can ...fluorophore-conjugated secondary antibodies specific to each isotype or species to easily discern the proteins...

  13. Visualizing Protein Turnover In Situ

    Type
    Blog Post
    ...protein turnover in situ in neurons. The classic approach for studying protein turnover is radioactive pulse-chase...immunocytochemistry is perhaps the most common approach. With a good and specific antibody and the proper...the proteins of interest are in close proximity to each other can the DNA strands take part in rolling circle...

  14. Viral Vectors 101: An Introduction to AAV

    Type
    Blog Post
    ...rates for AAVs are generally high enough for this approach to work, although it’s less effective in vivo ... of its biggest advantages. Specificity can be achieved using specialized promoters, Flp or Cre dependent...expression AAVs have to infect cells, hijack host cell machinery, and synthesize a second strand of DNA before ...

  15. Plasmids 101: The protein expression toolbox

    Type
    Blog Post
    ...introduced gene or over the native locus? Let’s explore each of the options together. Turn it off Turn it off...inducible expression (more on this below) will both achieve temporary repression of protein expression. Addgene...flavors of activators which can be used with this approach - read more about them on our CRISPRa blog! ...

  16. Antibodies 101: Fab Fragments

    Type
    Blog Post
    ...identical heavy chains and two identical light chains) each with a constant region and a variable region. The...regions, both through genetic and proteolytic approaches. This comes in handy when, for example, you want...product is a mini-Y with both Fab regions still attached to one another. Size: ~110 kDa. Fab' - These fragments...

  17. Lighting Up Cell Signaling with Photoswitchable Kinases

    Type
    Blog Post
    ...spatiotemporal resolution than pharmacological or genetic approaches, but only a few such methods exist and they only... used to turn “on” and “off” MEK1 activity by attaching two dimerizing photoswitchable Dronpa (dron= a...at sites homologous to the locations they were attached to in MEK. The photoswitchable versions of MEK1...

  18. Uncertainty about Labor Law Brings More Uncertainty to Postdoc Wages

    Type
    Blog Post
    ...biochemistry department at the University of Massachusetts Medical School. We are working with postdocs...these salary fluctuations, we are continuing to reach out to institutions with a statement indicating ...to hold. There are institutions in Boston - Massachusetts General Hospital, Mass Eye and Ear Infirmary...

  19. Plasmids 101: CcdB - The Toxic Key to Efficient Cloning

    Type
    Blog Post
    ...several methodologies currently available for approaching it. These include the traditional restriction...cloning technologies, as well as several others. Each method is unique and relies on specific components...   Figure 2: Image from http://teachline.ls.huji.ac.il/72682/tutorials/gateway/introduction.html...

  20. Addgene's Tips for Plasmid Quality Control

    Type
    Blog Post
    ...little extra and transfer 100 µl of glycerol stock to each of 2 96-well microplates (identical copies, see .... We then select forward and reverse primers for each sample with the help of DNA-editing software which...glycerol plate on dry ice to the sequencing provider. Each sequencing service will have different requirements...
Showing: 521 - 540 of 949 results