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We narrowed to 662 results for: MAL

Showing: 281 - 300 of 662 results

  1. Screening for Successful Genome Editing with Digital PCR

    Type
    Blog Post
    Published
    March 30, 2017, 2:30 p.m.
    ...cost-prohibitive for many labs, and are often impractical for small projects. Most researchers instead turn to so-called...frustrating to work with, time consuming, and minimally informative. In this blog post, we’ll introduce...one or no copies of the target DNA. Conventional thermal cycling is then performed to amplify the target...

  2. Viral Vectors 101: Gamma-Retroviral Packaging Systems

    Type
    Blog Post
    Published
    Aug. 6, 2024, 1:15 p.m.
    ...envelope, Phoenix-AMPHO (for the infection of mammalian cells.) Using this system, virus is produced in...the host cell cytoplasm, the viral components are small enough to enter the nucleus through the nuclear ...of retroviruses for efficient gene delivery to mammalian cells using 293T cell-based systems. Curr Protoc...

  3. Plasmids 101: Transformation, Transduction, Bacterial Conjugation, and Transfection

    Type
    Blog Post
    Published
    June 25, 2019, 12:54 p.m.
    ...infected bacterium can either exist as transient extrachromosomal DNA, like a plasmid, or it can integrate into...introduce foreign DNA into eukaryotic cells, like mammalian cell lines. This can be done with lentiviral and...chromosome and allows the donor bacterium to make a small “bridge” or sex pilus that attaches to the recipient...

  4. Viral Vectors 101: Viruses as Biological Tools

    Type
    Blog Post
    Published
    March 21, 2023, 1:15 p.m.
    ... There are four main types of viruses used in mammalian research: 𝛄-retrovirus, lentivirus, adenovirus...Mutagenesis Possible Adenovirus Remains Episomal (Little Risk of Insertional Mutagenesis) High ...Expression High Immunogenecity AAV Remains Episomal (Little Risk of Mutagenesis) Very Low Immunogenicity...

  5. Plasmids 101: Screens vs. Selections

    Type
    Blog Post
    Published
    Nov. 10, 2022, 2:15 p.m.
    ...or mutagenizing a population (typically cells or small organisms) and then assaying for a functional or...are genetic knockout mutations and treatment with small molecules/drugs. With the advent of CRISPR, genome-wide...be knocked out with CRISPR, the entirety of the small molecule realm is typically not feasible to test...

  6. Antibodies 101: ChIP

    Type
    Blog Post
    Published
    March 10, 2022, 3:25 p.m.
    ...therefore will require crosslinking, which uses formaldehyde or UV light to create strong but reversible ...Fragmenting  The next step is break the DNA into random, small pieces of roughly equal size, typically between ...Whichever method you choose, you’ll need to reserve a small aliquot, split into two parts to (1) run a DNA gel...

  7. Antibodies 101: Buffers, Storage, and Conjugates

    Type
    Blog Post
    Published
    Feb. 1, 2022, 2:15 p.m.
    ...going through small aliquots at a time, the -20°C is your best option, in aliquots no smaller than 10uL. ...guidelines below, you’ll likely be able to create a small, usable panel.  First, fluorophores vary in brightness...

  8. Viral Vectors 101: Inverted Terminal Repeats

    Type
    Blog Post
    Published
    Oct. 19, 2023, 1:15 p.m.
    ...scientists to express a gene of interest in a cell or animal model. AAV plasmids contain several important elements...deletion in the B or C domain of one ITR. These small deletions are not usually an issue as they are thought...enzymes cut at known sites within the ITR. However, small deletions and point mutations will go undetected...

  9. Protein Tagging with CRISPR/Cas9: A Conversation with Mendenhall and Myers

    Type
    Blog Post
    Published
    July 28, 2015, 2:30 p.m.
    ...things work correctly, the second plasmid inserts a small epitope tag called FLAG at the end of the transcription...beautiful ChIP-Seq results. The FLAG epitope is quite small—20 amino acids—so we don’t think it interferes with...challenges does this system help to solve? Myers: Only a small percentage of the proteins inside a cell have a ...

  10. Dos & Don'ts When Publishing a Scientific Manuscript

    Type
    Blog Post
    Published
    May 22, 2014, 4:02 p.m.
    ...identifier (a bit like a GenBank ID) to all antibodies, animal models, and software tools in the Materials and...contains all the necessary information to be of maximal use to the scientific communities. The ARRIVE guidelines... guidelines for the reporting of laboratory animals during in vivo research and the CARE guidelines for...

  11. Will You Be My Mentor? Finding and Asking for Mentoring Support

    Type
    Blog Post
    Published
    Feb. 4, 2014, 3:50 p.m.
    ...Broaden your network Take advantage of formal programs A formal mentoring program can be a great way to...year. Some click and some do not. These are all normal outcomes. If you have a broad range of mentors ...

  12. Visualizing Protein Turnover In Situ

    Type
    Blog Post
    Published
    Jan. 16, 2018, 3:20 p.m.
    ...regulation of protein abundance is essential for normal cell function, and, because I was studying a synaptic...protein, I wanted to measure it’s levels in its normal neuronal context. In other words, I needed a method...microscopy techniques - Which is best for me? Learn how small molecule fluorophores can be used for cell labeling...

  13. Troubleshooting Your Plasmid Cloning Experiment

    Type
    Blog Post
    Published
    Sept. 24, 2019, 12:55 p.m.
    ...new plates with a fresh stock of antibiotic. Many small colonies surrounding big colonies. You probably ...left your plate for too long in the incubator. The small colonies are called satellite colonies and they ...screening Pick a few colonies (5-10) and grow them in a small culture volume (2 or 3 ml) containing the corresponding...

  14. CRISPR 101: Cas9 vs. The Other Cas(s)

    Type
    Blog Post
    Published
    Dec. 8, 2022, 2:15 p.m.
    ...therapeutic promise due to the fact that it is smaller than the traditional SpCas9 enzyme (about a third...sequence Similar to Cas12, it is also significantly smaller in size than traditional Cas9. Another similarity...RNA cleavage and is significantly less toxic to mammalian cells than Cas13 (likely due to the lack of collateral...

  15. Giving gRNAs a Facelift - Synthetic and Beyond

    Type
    Blog Post
    Published
    Oct. 20, 2022, 1:15 p.m.
    ...usually 1-3 of the first 3 bps are modified) for optimal stability and efficacy.  Base modifications What...are slightly more efficient and less toxic to mammalian cells (Allen, et al., Frontiers in Genome Editing...RNAs to Enhance CRISPR Genome Editing Systems in Mammalian Cells. Frontiers in Genome Editing, 28 (2021)....

  16. Pairing CombiGEM and CRISPR for Combinatorial Genetic Screening

    Type
    Blog Post
    Published
    April 12, 2016, 2:30 p.m.
    ...in gRNA design necessary to achieve maximal on-target and minimal off-target activities have proven themselves.... "Genome-wide recessive genetic screening in mammalian cells with a lentiviral CRISPR-guide RNA library...

  17. Teaching an Old DOG New Tricks: Controlling Protein Activity with GFP

    Type
    Blog Post
    Published
    Nov. 24, 2015, 3:30 p.m.
    ..., consist of a single heavy chain, and they are small and stable inside cells. Nanobodies binding GFP ...and since cells infected with AAV-Cre-DOG retain normal neuronal function, this system may make optogenetics...developed soon to allow genome modification only in a small subset of cells? Other proteins may also be utliized...

  18. Overcoming the Challenges of Lentiviral Production

    Type
    Blog Post
    Published
    Aug. 7, 2018, 12:31 p.m.
    ...(i.e. polyethyleneimine, calcium phosphate or liposomal methods), the volume of the reaction, and the ...produces high titer virus for transfer plasmids with small inserts (such as CRISPR guides) while a single 72h...viral particles allowing you to concentrate in a smaller volume. Consequently, you will end up with a considerably...

  19. Plasmids for Endogenous Gene Tagging in Human Cells

    Type
    Blog Post
    Published
    April 6, 2017, 1:02 p.m.
    ...protein, which can both interfere with a cell’s normal function and result in a ubiquitous background ...necessary to ensure the edited cells are behaving normally can be prohibitively expensive for many labs. ...Tagging collection includes systems for tagging mammalian cell lines, Drosophila cells, and C. elegans. ...

  20. Antibodies 101: Stripping and Reprobing Western Blots

    Type
    Blog Post
    Published
    Aug. 1, 2024, 1:15 p.m.
    ...western blot Run your gel and membrane transfer as normal. Immediately after transfer, dry the membrane to...first. Then continue on with your western blot as normal! Stripping your membrane Alright, so you’ve dried...antibody, you’re ready to repeat your western blot as normal. Be sure to rinse away the secondary antibody you...
Showing: 281 - 300 of 662 results