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We narrowed to 815 results for: MAL

Showing: 381 - 400 of 815 results

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

    Type
    Blog Post
    ...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...

  2. Viral Vectors 101: Viruses as Biological Tools

    Type
    Blog Post
    ... 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...

  3. Viral Vectors 101: Gamma-Retroviral Packaging Systems

    Type
    Blog Post
    ...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...

  4. Plasmids 101: Screens vs. Selections

    Type
    Blog Post
    ...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...

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

    Type
    Blog Post
    ...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 ...

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

    Type
    Blog Post
    ...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...

  8. Giving gRNAs a Facelift - Synthetic and Beyond

    Type
    Blog Post
    ...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)....

  9. Troubleshooting Your Plasmid Cloning Experiment

    Type
    Blog Post
    ...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...

  10. Visualizing Protein Turnover In Situ

    Type
    Blog Post
    ...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...

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

    Type
    Blog Post
    ...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...

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

    Type
    Blog Post
    ..., 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...

  14. Plasmids 101: In-house NGS Plasmid Quality Control Now at Addgene

    Type
    Blog Post
    ...on each and every isolated plasmid sample and normalize each to fall within a 10 fold range. Plasmids ...Since NGS assembles a larger sequence from many smaller sequences, regions with repeats may not assemble...samples are filled into the same process as our normal Complete Plasmid Sequencing (CPS) workflow and ...

  15. Plasmids for Endogenous Gene Tagging in Human Cells

    Type
    Blog Post
    ...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. ...

  16. Overcoming the Challenges of Lentiviral Production

    Type
    Blog Post
    ...(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...

  17. Viral Vectors 101: Calcium Sensors

    Type
    Blog Post
    ...neuron-specific expression (hSyn promoter) and pan-mammalian cell expression (CAG promoter). An improved version...neuron-specific expression in both zebrafish and mammalian systems. The GCaMP6, jGCaMP7, and jGCaMP8 sensors...neuron-specific, astrocyte-specific (GCaMP6 only) or mammalian cell expression, and compatible with constitutive...

  18. Viral Vectors 101: Chemogenetics

    Type
    Blog Post
    ... to probe cells and circuits without disrupting normal brain function. Chemogenetics has become a popular...successfully reach target cells while resulting in minimal off target effects. For example, if your receptors...endogenous gene expression, which can lead to changes in normal cell physiology and function (Saloman, JL, et al...

  19. Antibodies 101: Stripping and Reprobing Western Blots

    Type
    Blog Post
    ...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...

  20. Progress Towards a PAM-Free CRISPR

    Type
    Blog Post
    ...desired edit to still occur if the cut site is sub-optimal? As with everything in biology, the answer is always...possible to "get lucky" and find a great gRNA with a proximal PAM right where you need it, the odds of this ...genome! Want to get your hands on SpRY? Find a mammalian plasmid here and edit on! References and Resources...
Showing: 381 - 400 of 815 results