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  1. Visualizing Genomic Loci with CRISPR-Sirius

    Type
    Blog Post
    ... arrangements may affect nuclear processes. The latest of these visualization tools is CRISPR-Sirius, ...specific loci based on the guide RNA sequence. A year later, they developed CRISPRainbow (Ma et al., 2016), ...chromosome-specific loci in the genome and the guides are relatively unstable.   Figure 1: CRISPRainbow labeling... proteins. (B) CRISPR-Sirius used to visualize relative locations of the subtelomeric region of the pericentromeric...
  2. 5 Great Apps for Lab Life

    Type
    Blog Post
    ...tried to find an app that could help me with calculations. This is how I came across the Gibco Cell Culture...culture and include a counter, lab timer, dilution calculator, and a couple of other nice features. This app...sequence for your enzyme of interest. Lastly, the Tm calculator helps you set up an optimal PCR depending on ...apps: Findings Lab Notebook Cell Culture Split Calculator by DRDP Restriction Enzyme Tool by Promega Material...
  3. Sweating the Small Stuff: Details in the Lab

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    Blog Post
    ... to glance at as you work. Draw your plate maps onto your plates, alongside your physical copy. If you...reminders on a sticky note   Visual Planning  Plate maps, pre-labeling tubes, and even drawing out experimental...example, setting up your pipette tips to match your plate layout can help you track samples as you add them...
  4. GPCRs: How Do They Work and How Do We Study Them?

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    Blog Post
    ... in the nucleus. The Roth lab showed that this platform accurately identified 120 known GPCR targets demonstrating..., the receptor fused to APEX2 catalyzes the biotinylation of proteins near the GPCR. These proteins are... GPCRs for chemogenetic studies to probe the relationship between neurons and specific behaviors. Chemogenetics...DREADDs are ideal for chemogenetics as they are relatively insensitive to their endogenous ligand, acetylcholine...
  5. Viral Vectors 101: Biosensors

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    Blog Post
    ... was the molecule of interest (analyte), and accumulation of ATP activated GFP expression (receptor), ..., then the transducer could be a microscope, plate reader, or any other instrument which measured GFP ...advantages over other mechanisms. For one, many viral platforms, such as AAV or retrovirus, can either permanently...Fluorescent Biosensors for Neurotransmission and Neuromodulation: Engineering and Applications. Frontiers in...
  6. Viral Vectors 101: Viral Applications

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    Blog Post
    ...of your library cargo to isolate the effects of each variable being manipulated. Thus, the adjustable nature...expression control Researchers are always wanting to manipulate gene expression – turn it up, turn it down, knock...PMC2075523. Bulcha, J.T., et al. Viral vector platforms within the gene therapy landscape. Sig Transduct...
  7. Antibodies 101: ChIP

    Type
    Blog Post
    ...of interest (4) isolated your antibody:protein:DNA complexes and finally (5) isolated the DNA from your...DNA are no longer bound, and the DNA can then be isolated and studied through a variety of downstream methods...downstream analysis via any methods suitable for isolated and purified DNA, such as sequencing or qPCR. ...
  8. Multiplex Genome Editing with CRISPR-Cpf1

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    Blog Post
    ...multiplexing vectors are larger because they require regulatory sequences to allow for expression of multiple...example of how this works. Make sure to order 5’ phosphorylated oligos or treat with T4 PNK. Delivery options... are easier to use across multiple expression platforms: transfection, lentiviral transduction, and AAV...Multiplex gene editing with CRISPR-Cpf1 is one of the latest developments in CRISPR technologies.  It’s a simple...
  9. Form Your Own Peer Mentoring Group: A How-To Guide for Scientists

    Type
    Blog Post
    ...participants in skills useful for future mentoring relationships.   If you would like to start a group mentoring...for the time commitment of a formal mentoring relationship or there aren’t enough senior mentors available...ideas on what to do once you are in a mentoring relationship.  Celebrate your successes and support each... come. Are you involved in Peer Mentoring relationships? Comment below with ideas from your own mentoring...
  10. Plasmids 101: The protein expression toolbox

    Type
    Blog Post
    ...mechanism? Where is the control happening? Is the regulation of an exogenously introduced gene or over the...generate a frameshift mutation that will permanently ablate protein expression. If you need the ability to...different amounts of tetracycline. Tetracycline regulation is reversible (when the drug is removed expression... These control systems function at the post-translational level, unlike most of the other mechanisms described...
  11. Antibodies 101: Fab Fragments

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    Blog Post
    ...The modularity of antibodies means that it is relatively easy to physically divide up the different regions...you to use just a fragment of the antibody in isolation from another portion. For example, Fab fragments...but not the Fc region (more on when that might be later). Find scFv plasmids at Addgene Depending on how...fragments intended for use as primary antibodies are relatively uncommon. Though, if you want to use Fab fragments...
  12. Giving gRNAs a Facelift - Synthetic and Beyond

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    Blog Post
    ... with post translational modifications. One of the most common of these is 2’-O-methylation, where a methyl...once you’ve chosen a gRNA and modification(s) Congratulations! You’ve picked your perfect guide; now check...Spatiotemporally Controlled CRSIPR-Cas-based Genomic Regulation. ACS 6, 5, 695-703 (2020). https://pubs.acs.org...
  13. The Breast Cancer Microenvironment: A Tumor’s Backstage Team

    Type
    Blog Post
    ... TAK 1, an enzyme that controls apoptosis and regulates transcription. Without TKA1, cells proliferate...cancer as a broad phenomenon and not only as an isolated disease. While the tumor may seem to take center...microRNA transfer: A mechanism of environmental modulation of hepatocellular cancer cell growth." Hepatology...resources on the Addgene blog Read our cancer related blog posts Learn about our cancer collection pages...
  14. Choosing the B(right)est Fluorescent Protein: Photostability

    Type
    Blog Post
    ... not linearly related to power, the conclusions reached at high power may not translate to real applications...widefield imaging fluorophores are excited with relatively low light levels (for a prolonged time). Other...application (Goedhart et al, 2012). The third issue relates to the environment of the fluorescent protein. ...
  15. Pairing CombiGEM and CRISPR for Combinatorial Genetic Screening

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    Blog Post
    ... that underlie complex regulation in biological systems. The CombiGEM platform provides a means to create.... CombiGEM-CRISPR: The marriage of two simple platforms CRISPR has revolutionized how we decode the genome...Wong et al., 2016). With CombiGEM and CRISPR platforms now being integrated, we look forward to the realization...
  16. Addgene's Tips for Plasmid Quality Control

    Type
    Blog Post
    ...samples through our lab using barcoded tubes and plates. The critical pieces of information for a given...100 µl of glycerol stock to each of 2 96-well microplates (identical copies, see image above). We then ... database of over 2000 primers. Two 96-well PCR plates are prepared with the primer stocks for forward...shipped along with their corresponding glycerol plate on dry ice to the sequencing provider. Each sequencing...
  17. Pathways Over Time Plasmids Engage Students in Functional Genomics Research

    Type
    Blog Post
    ...acquired divergent functions due to mutations accumulated over time? Undergraduates can help to answer ...If the phenotype is corrected, students can be relatively confident that the gene from the test species...the S. cerevisiae met deletion strains and then isolate transformed strains on media that lack uracil, ... MET genes to grow.) Students then use replica plating to determine if expression of MET/Met genes from...
  18. Hot Plasmids - October 2020

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    Blog Post
    ...glycosyltransferase genes and other proteins related to cellular glycosylation. Get the library at Addgene CasRx...requires the much less efficient double stranded templates, rather than the more efficient single-stranded...cellular protein. They used CRISPR with a ssODN template to introduce the small wrmScarlet11 which is less...
  19. Fluorescent Proteins 101: Introduction to FRET

    Type
    Blog Post
    ...donor-acceptor pair and a change in FRET efficiency correlates with a change in the distance and/or orientation...available. Fortunately, several excellent reviews have collated data from various papers to make the decision ...conformation of the probe. More on these biosensors later on). Experiments using intermolecular FRET probes...simultaneously would further increase the amount of correlated information available from a cellular process...
  20. Nanoblades: Tiny CRISPR Ninjas for Genome Editing Difficult Cells

    Type
    Blog Post
    ...pluripotency marker expression and response to LPS stimulation, respectively. Primary cell type Gene targeted... a fluorophore or tag. To utilize HDR, a donor template containing the desired sequence flank by homology...Nanoblades which carries a gRNA, Cas9, and a HDR donor template DNA can be generated by incubating Nanoblade particles...facilitated HDR with either a ssDNA or dsDNA donor template. In vivo editing of mouse zygotes and mice with...
Showing: 521 - 540 of 955 results