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We narrowed to 297 results for: IRES

Showing: 61 - 80 of 297 results

  1. Plasmids 101: The Promoter Region – Let's Go!

    Type
    Blog Post
    ...binding sites, commonly used in Drosophila Specific Requires the presence of Gal4 gene to activate promoter...Promoter from T7 bacteriophage Constitutive, but requires T7 RNA polymerase. When used for in vitro transcription... Negligible basal expression when not induced. Requires T7 RNA polymerase, which is also controlled by...Promoter from Sp6 bacteriophage Constitutive, but requires SP6 RNA polymerase. SP6 polymerase has a high ...Promoter from T3 bacteriophage Constitutive, but requires T3 RNA polymerase When used for in vitro transcription...

  2. CUT&RUN: An Improved Method for Studying Protein-DNA Interactions

    Type
    Blog Post
    ...materials. Problems with Chip-seq ChIP-seq typically requires a large amount of input material, cells or tissue...experiment can be a major technical hurdle.   ChIP-seq requires  cross-linking during an initial fixation step...wing as a model. A typical ChIP-Seq experiment requires around 1,000 wings. Collecting that much tissue...controlled by the addition of calcium, which MNase requires for its nuclease activity and is chelated from...

  3. Advice for Scientists Starting a Lab

    Type
    Blog Post
    ...select first hires who are well-suited, productive, and fit well with your goals. These first hires will set...set the tone and atmosphere that future hires will look up to." She also recommends to seek out advice...lab and selectively picking the first couple of hires to build a strong, collaborative team. And take ...

  4. Finding and Joining Your Dream Lab

    Type
    Blog Post
    ...three lab heads how they go about selecting new hires. This is followed by some perhaps surprising, yet...scientists for their labs. Tom Ellis selects new hires from a pool of CVs collected from throughout the...are dedicated and interested in science. “Postdoc hires are simpler because of their documented history ...with their interests.” George Church selects new hires initially based on those who notice and embrace ...

  6. Common Injection Routes in Mice

    Type
    Blog Post
    ...seem like the easier of the two, but it actually requires more technical skill to place the needle in the...the vertebrae, so hitting the correct location requires a lot of skill and anatomical knowledge. Intrathecal... the lungs. It is an advanced technique, as it requires intubation of the mice, which can be tricky given... intranasal is easier, less invasive, and only requires a pipette. Since the substances are administered...

  7. Production of Virus in Insect Versus Mammalian Cells

    Type
    Blog Post
    ...preps, lentiviral titration requires an additional couple of days, as it requires transduction of more cells...companies. Production of both AAV and lentivirus requires three plasmids — a transfer plasmid containing...baculovirus plasmid. Production of baculovirus requires only two plasmids — your transfer plasmid and ...

  8. Tips for a 1st time AAV user (by a Rookie AAV user)

    Type
    Blog Post
    ...AAV replication uses the host’s polymerase, but requires Rep proteins to process a double-stranded intermediate...AAV than I think I need. Unlike lentivirus, AAV requires a higher number of viral particles for efficient...several hours on a heated stir plate to dissolve and requires monitoring so that the solution doesn’t get too...AAV for my cells of interest. Every batch of AAV requires its own MOI optimization to account for batch-...

  9. Don’t FRET: Bimolecular Fluorescence Complementation Makes Visualizing Protein-Protein Interactions Easy

    Type
    Blog Post
    ... of hard-to-achieve factors. For example, FRET requires that fusion proteins be in close proximity and...FRET, as the overexpression of fusion proteins it requires can lead to aberrant subcellular localization ... construction Fusing FP fragments to your PIPs requires careful selection of FP fragments and, preferably...information, constructing appropriate controls may requires that you empirically test a number of mutants ...

  10. Technical Design of a Western Blot

    Type
    Blog Post
    ...conjugate, and changing your reporter molecule requires either re-conjugating or re-purchasing your primary...most protein from your gel to your membrane. It requires anywhere from sixty minutes to overnight to run... can be done.  Note that each type of transfer requires different equipment. Wet transfers can be done...compatible with chemiluminescence reporters and requires access to a darkroom and a developer. Film is ...exposures for a blot with varying signals on it. It requires long exposure times (up to thirty minutes!) to...

  11. Harnessing Bacterial Toxins for Allelic Exchange

    Type
    Blog Post
    ...mutations. Ultimately, successful allelic exchange requires two sequential homologous recombination events...events, but because these events are rare, each requires selection. These two steps are (Figure 1): The ...and excision of the vector backbone. This step requires “counter-selection,” meaning that the selection...generate precise, unmarked mutants, but it frequently requires considerable optimization to ensure strong counter-selection...

  12. Fluorescent Proteins 101: When GFP lets you down

    Type
    Blog Post
    ...myristoylation requires an N-terminal consensus sequence (MG-) and prenylation requires a C-terminal consensus... 2002). GFP needs oxygen Chromophore formation requires a fully folded beta-barrel structure, followed...chromophore (Tsien, 1998). Importantly, this reaction requires molecular oxygen and as a consequence GFP remains...

  13. Genetic Code Expansion

    Type
    Collection
    ...182653 pcDNA3.1(+)_U6 tRNAPyl_CMV NESPylRS(AF)_IRES_eRF1(E55D)-HA Y306A/Y384F (AF) pyrrolysine (Pyl) tRNA...

  14. Delivery Methods for Generating iPSCs

    Type
    Blog Post
    ...promoters for transcriptional regulation, so it requires a different approach for regulating transgene ...Generating iPSCs with plasmid-based expression requires serial transfection of 1 or 2 plasmids that express... the gold standard instead. Teratoma formation requires injecting iPSCs into immunodeficient mice and ...suited for these situations. The second scenario requires higher levels of safety and as little chance of...

  15. Antibodies 101: Affinity Tags

    Type
    Blog Post
    ...must be performed after primary purification and requires a second clean up step to remove the cleaved tag...Also, purifying GST by affinity chromatography requires proper folding of the tag, which prevents insoluble.... Mammalian purification is still possible but requires optimization and often more stringent washing....

  16. Pushing the Limits of DNA Assembly

    Type
    Blog Post
    ... endeavor, NEBuilder requires upfront planning. This cloning methodology requires that the fragments to...simplify this process by using NEBuilder which requires only a single-stranded synthetic oligo to generate...

  17. Visualizing Translation at the Single Molecule Level

    Type
    Blog Post
    ...frequency. Although highly useful, this technique requires extensive sample processing to avoid disrupting...appear red. To enable this color switching, TRICK requires the following components: a GFP protein that ...caveats. In both cases, creating a reporter mRNA requires inserting multiple hairpin sequences into the ...

  18. Choosing Your Fluorescent Proteins for Multi-Color Imaging

    Type
    Blog Post
    ...simultaneously. To do so with fluorescent protein labels requires multiple fluorescent proteins whose excitation... means that the choice of fluorescent proteins requires some thought. The first step in choosing fluorescent...fluorescent proteins can be separated from each other requires knowledge of their excitation and emission spectra...

  20. RANbodies: Reporter Nanobody Fusions

    Type
    Blog Post
    ...are a non-renewable reagent - Their production requires animal sacrifice and because they are an animal...secondary Less sensitive than HRP-RANbody staining Requires secondary antibody staining HA Tagged GFP, RFP...secondary   Less sensitive than HRP-RANbody staining Requires secondary antibody staining GFP, RFP, H2A2B ...
Showing: 61 - 80 of 297 results