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We narrowed to 892 results for: CAN

Showing: 461 - 470 of 892 results

  1. New Optimized Genome-wide CRISPRko, CRISPRi, and CRISPRa Libraries

    Type
    Blog Post
    Published
    Oct. 4, 2018, 12:44 p.m.
    ...efficiently investigate gene function. CRISPR libraries can be used to knock out, inhibit or activate target ...function while nuclease-deactivated Cas9 (dCas9) can be tethered to a repressor (CRISPRi) or an activation... are subject to an increase in dsDNA breaks that can cause cell death. Thus, CRISPR inhibition (CRISPRi...In addition to inhibiting gene expression, dCas9 can also be used for transcriptional activation. Activation...genome-wide gain of function screens. These screens can help reveal the function of lowly expressed genes...Tanenbaum et al., 2014). The sgRNA or tracrRNA sequences can also be modified to include RNA domains that recruit... screen, implying that ORF and CRISPRa libraries can be used in complement to study gene function. Find...

  2. Plasmids 101: Control Plasmids

    Type
    Blog Post
    Published
    April 9, 2015, 3:29 p.m.
    ...investigator needs a standard against which the results can be compared; results from an experiment lacking the...on to learn more about the various controls that can be used for plasmid-based experiments. What are control...expression level of Gene X in the untreated cells—but can we be certain? Do we know whether the plasmid delivered...the baseline/standard against which other samples can be compared. The Empty Vector Control (mentioned ...and biological. In general, technical replicates can be thought of as "plate controls". They are NOT independent...Conversely, biological replicates ARE independent and can be thought of as "reproducibility controls". Biological...successfully transfected our cells (Figure 3), we can continue with our experiment, incorporating additional...

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

    Type
    Blog Post
    Published
    April 3, 2014, 8:05 p.m.
    ...RNAs. To gain this specificity, the eukaryotic RNAP can recognize and bind to specific promoter elements....cytomegalovirus Constitutive  May contain an enhancer region. Can be silenced in some cell types. EF1a General expression...and several tetracycline operators. Transcription can be turned on or off depending on what tet transactivator...Inducible with galactose; repressible with glucose  Can be used independently or together. Regulated by GAL4...polymerase, which is also controlled by lac operator. Can be induced by IPTG. Commonly found in pET vectors...in the absense of lac repressor (lacI or lacIq). Can be induced by IPTG or lactose. Leaky promoter with...gene expression  Promoter from bacteriophage lambda Can be temperature regulatable  Often paired with the...

  4. When Fidelity Matters: A frank discussion about ligase fidelity

    Type
    Blog Post
    Published
    March 6, 2018, 1:49 p.m.
    ...mismatches. However, some ligases can ligate mismatches to a significant degree, and very active ligases...dependent DNA ligase from a thermostable bacterium that can survive high temperatures (up to 95 °C) and is active...right ligase, conditions and suitable probes, LDR can reliably detect single-base differences. The original...visualized the ligation product in a gel, but we can now use fluorophore- quencher pairs or qPCR-based...ligases, such as T4 DNA Ligase, can ligate nicks containing one or more mismatches near the ligation junction...This “peeling back” of the 5 ́-phosphorylated base can be observed in the crystal structures of several ...26). Despite this higher fidelity, Taq DNA Ligase can still detectably ligate many T:G, T:T, and A:C mismatches...

  5. Fluorescent Proteins 101: Monitoring Cell Mobility Using Fluorescent Proteins

    Type
    Blog Post
    Published
    Aug. 15, 2017, 1:24 p.m.
    ... they can scan for any signs of danger. In this post we’ll focus on how fluorescent proteins can and have...incremental change of the excitation wavelength can significantly affect emission intensities from fluorescent...coming from this volume. The consequence is that you can detect signal in thick specimens. See the video below...lymph node after infection with S. aureus (red). You can then see neutrophils swarming around S. aureus and...interactions. Even if virtually all fluorescent proteins can be used in your experiments, experience has shown...excitation wavelength (Salomonnson et al. 2012). They can also be excited efficiently at wavelengths between...the the Cre/lox recombination system. A researcher can flank fluorescent proteins in a plasmid with loxP...

  6. Tagging Optogenetics and Chemogenetics Receptors: Fluorescent Proteins and Other Options

    Type
    Blog Post
    Published
    May 11, 2021, 1:15 p.m.
    ...or post-mortem experiments. Fluorescent proteins can also be identified using antibodies and immunohistochemistry... always be the best option. Fluorescent proteins can interfere with fused protein function Under some ...some circumstances, the fused fluorescent proteins can be a roadblock, rather than an aid, in the expression...., 2013). We found that the fluorescent proteins can, indeed, interfere with the correct function of the...learned from these studies is that the tag protein can impact the expression of the transgene of interest...provided evidence that some fluorescent proteins can be, in some circumstances, detrimental to the expression...chemogenetic receptors used in these techniques cannot be directly visualized with fluorescence or immunohistochemistry...

  7. Antibodies 101: Fab Fragments

    Type
    Blog Post
    Published
    Sept. 5, 2024, 1:15 p.m.
    ...assembled, the four chains make a Y-shaped protein that can be divided into the Fragment Crystallizable region...allows them to get into samples more efficiently and can also improve localization accuracy when used in super-resolution... to use Fab fragments as primary antibodies, you can make your own antibody fragments as needed.  In contrast...complex into the sample. Alternatively, Fab fragments can be used as a block in similar scenarios. By applying... here? Many of the Fab fragments described above can be used for similar applications and have similar...interchangeable, but the extra bit of heavy chain on a Fab’ can act as a convenient site for conjugation.  Finally...

  8. Viral Vectors 101: Calcium Sensors

    Type
    Blog Post
    Published
    Aug. 10, 2023, 1 p.m.
    ...Unlike chemical indicators, these DNA constructs can be delivered to intact tissues and organs noninvasively... selecting a sensor to use for your studies, you can start by asking ‘What is the biological phenomena...concentrations in different subcellular compartments can vary greatly. Additionally, the calcium changes you...interference, and comparison between experiments can be more difficult due to variance in sensor expression...GEX or blue/green CEPIA) or through FRET sensors, can reduce these unwanted variables. Twitch is a FRET-based... By employing various AAV serotypes, researchers can precisely direct the calcium sensor to specific tissues...delivered using viral vectors! One of the most significant advantages of viral delivery is the potential...

  9. Transferable Skills Guide: Public Speaking

    Type
    Blog Post
    Published
    April 20, 2023, 4 p.m.
    ...make a strategy that can help. Even something as simple as telling yourself, “I can stop and check my notes...Most of the time, that’s enough–but if it’s not, I can always take more time to check my notes!  Know your... speaking, Murphy’s law definitely applies: what can go wrong, will go wrong. The trash removal service...to,” or “It is okay if it doesn’t go perfectly,” can help. (And do remember to breathe!)   Public speaking... a chance to comfortably engage with you. If you can give a seminar, lead a journal club, or present at.... The whole process took just under a minute. I can't say the journal club was a rousing success–the response...

  10. Transferable Skills Guide: Cross-team Communication

    Type
    Blog Post
    Published
    Nov. 21, 2019, 1:53 p.m.
    ...buy-in from all of Addgene’s teams. This solution can easily be implemented in an academic lab setting,...ambiguous terms. Defining words and applying annotation can be particularly useful in academic labs when creating...Clarify: Asking for clarification during a meeting can help get everyone on the same page. Others in the...meeting might also have the same question and asking can help the team identify possible points of confusion... done. But if you implement it well, here’s what can happen: More effective meetings. When everyone at...people aren’t listening to you or understanding you can be incredibly frustrating. Clarifying the words —...
Showing: 461 - 470 of 892 results