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We narrowed to 289 results for: poly

Showing: 41 - 60 of 289 results

  1. 5 Tips for Troubleshooting Viral Transductions

    Type
    Blog Post
    ...negatively-charged membranes (Davis et al., 2002). Polybrene is one such cationic reagent that is commonly ...efficiency by 10-fold (Davis et al., 2002). However, polybrene is highly sensitive to freeze-thaw cycles and ...be stored in single-use aliquots. Never use a polybrene solution that has been thawed more than once. ...in cells that are sensitive to the toxicity of polybrene (e.g., hematopoietic or primary cells), and has...., Jeffrey R. Morgan, and Martin L. Yarmush. "Polybrene increases retrovirus gene transfer efficiency ...

  2. When is a Monomer not a Monomer? The Top Three Ways Your Favorite Fluorescent Protein Oligomerizes in Cells

    Type
    Blog Post
    ...sufficient to seed polymer formation as fusion protein concentration increases. The polymerized fusion proteins...dimerizing FP has potential to form large stable polymers of dimers. 3) FRET biosensors that contain two...dimerization coupled with POI dimerization can lead to polymerization of the fusion protein. A dimeric fusion protein...resulting in false positives [2, 4]. Solutions to the polymer problem There are some simple solutions available...

  3. MXS Chaining

    Type
    Blog Post
    ...not suited to generate polycistronic containing multiple identical polyadenylation signals or promoters ...fluorescent proteins, promoters, enhancers, polyadenylation signals, inducible gene-expression sequences...promoter (to drive high-level expression) and a polyA signal (to terminate transcription) and the four...constructs were then combined to create a single polycistronic 15 kb insert. The final construct was then introduced...

  4. The Materials Science of Optogenetics Experiments

    Type
    Blog Post
    ... impossible to interpret. Polishing the fiber optics with diamond polishing paper of sequentially increasing... second key point; the fiber optics need to be polished to significant smoothness to prevent drastic loss...its output with a light meter before and after polishing. Efficiency of the implant should be at least ... output. The same considerations apply to the polishing and preparation of the patch cable (although you...

  5. Changing Labor Laws Bring Increased Postdoc Wages

    Type
    Blog Post
    ... how much they get paid. Institutional salary policies are one indicatator of how salaries may changes... 2014, 89% of institutions had a minimum salary policy: of those, 51% of institutions set their postdoctoral...National Postdoctoral Association Institutional Policy Report 2014). However, ~30% set their minima lower...lower. Furthermore, 7% did not enforce these policies. It is unclear how many postdocs are at each of ...only is this in direct contradiction to explicit policies set up other institutions (like University of ...

  6. Antibodies 101: Introduction to Antibodies

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    Blog Post
    ...target protein, so they are referred to as polyclonal. Polyclonal antibodies have the advantage of more robustly...find more details about polyclonal antibodies on our Antibodies 101: Polyclonal Antibodies blog post. Monoclonal...advantages and disadvantages compared to each other. Polyclonal antibodies These antibodies are derived from ...

  7. CRISPR 101: Multiplex Expression of gRNAs

    Type
    Blog Post
    ...express fewer than four gRNAs, you’ll clone in a polyT-termination sequence for each unused promoter. This...transcript You can also multiplex gRNAs via a polycistronic transcript. Rather than being transcribed from...option described below, plasmids for making polycistronic gRNAs are also available from the Yang lab for...Comparison of Multiplex Strategies including Standard PolIII-gRNA cassette, Csy4-cleavable cassette, and PTG...Plasmids: These plasmids are similar to the Csy4 polycistronic system described above, except that they use...gRNAs are flanked by glycine tRNAs to create polycistronic glycine tRNA-gRNA (PTG) constructs. Eukaryotic...

  8. Bringing Sustainable Practices to the Lab: Recycling

    Type
    Blog Post
    ... boxes Some vendors take back their expanded polystyrene (a.k.a. Styrofoam or cooler boxes). We get these...receiving temperature-controlled reagents. Expanded polystyrene will likely end up in a landfill, but at least...with CO2 emissions. NEB takes back expanded polystyrene boxes for free (in the U.S.). Their cooler boxes... 1976! Millipore Sigma takes back expanded polystyrene boxes for free (in the U.S.). After you receive...

  9. FastCloning: A Newer, Simpler Cloning Technique

    Type
    Blog Post
    ... excessive PCR cycles or low fidelity Taq DNA polymerase, the construction of specific sequences for base...believed that the exonuclease activity of the polymerase generates sticky ends in the PCR reactions. These...any subsequent steps. Using high fidelity DNA polymerases, I have never had any mutations introduced into...efficiency is highest when using high fidelity DNA polymerases like phusion. An 18-cycle PCR leads to the most...

  10. Viral Vectors 101: Parts of the AAV Transfer Plasmid

    Type
    Blog Post
    ...just upstream of the final ITR.  polyA  The polyadenylation or polyA signal aids in the nuclear export...translation, and promotes RNA transcript longevity. The polyA is located at the end of the AAV genome, just upstream...

  11. Plasmid Cloning by PCR

    Type
    Blog Post
    ...use a high fidelity polymerase to minimize mutations. The fidelity of the polymerase becomes more important... million bp depending on the polymerase used. No matter which polymerase you use, it is important that...

  12. Antibodies 101: Monoclonal Antibodies

    Type
    Blog Post
    ... be shared across other proteins. Conversely, polyclonal antibodies - which are a heterogenous mixture...antibodies, their cost is higher than that of polyclonal antibodies. For example, it takes longer to develop...than it does to immunize an animal and recover polyclonal antibodies (Lipman et al., 2005). This is reflected...Trudel LJ, Weis-Garcia F (2005) Monoclonal Versus Polyclonal Antibodies: Distinguishing Characteristics, Applications...

  13. Delivery Methods for Generating iPSCs

    Type
    Blog Post
    ...cloning capacity of ~8 kb, so either a single polycistronic vector or multiple single-gene vectors can be...retroviral vectors. This makes it possible to use a polycistronic cassette that expresses multiple reprogramming...factors can be delivered either as a single polycistronic transgene or with four different adenoviruses... ~14.5kb minicircle expressing a 2A peptide polycistronic cassette comprised of OCT4, SOX2, LIN28, NANOG...help mediate their transduction, such as the polyarginine peptide described in Zhou et al and Kim et ...

  14. CRISPR/Cas9 FAQs Answered!

    Type
    Blog Post
    ... and surveyor. Q15: I used DNA polymerase Takara Ex Taq™ DNA Polymerase for my genomic PCR, but couldn't...In our hands, Herculase II Fusion polymerase or Kapa Hifi Polymerase work very well. Some groups have ...your HR by doing Restriction Fragment Length Polymorphism (RFLP) (see Figure 4 of the Cong et. al (2013...

  15. Plasmid-based Recombinant Monoclonal Antibodies: What They Are and Why You Should Be Excited About Them

    Type
    Blog Post
    ... Traditionally, they have been made as either polyclonal antibodies or monoclonal hybridomas, but those...those techniques have several drawbacks. Polyclonal antibodies produced in animals may be variable among...& Plückthun, 2015a).   Figure 1: (Top) Polyclonal antibodies are produced in animals and consist...Recombinant antibodies offer numerous advantages over polyclonal antibodies and traditional hybridomas.  First...

  16. Deep Dive: Fixing and Permeabilizing for Immunofluorescence

    Type
    Blog Post
    ...slow the polymerization of formaldehyde.  Note: Formaldehyde in solution will naturally polymerize and lose...paraformaldehyde (PFA) as the fixative. PFA is a polymer of formaldehyde that, on its own, is not a fixative...PFA is dissolved in solution and heated, it depolymerizes, producing formaldehyde. Often people will call...

  17. Getting to Know Expansion Microscopy

    Type
    Blog Post
    ...biomolecules to the gel and add gel components to polymerize. Step 3: digest the sample to allow for even ...components penetrate the sample, how well the gel polymerizes, and the level of digestion. All of these factors...of epitopes during sample anchoring and gel polymerization - issues that can be minimized through tweaks...tweaks to these steps. But even under ideal polymerization and digestion conditions, loss of signal often...

  18. Simplify Cloning with in vivo Assembly

    Type
    Blog Post
    ... both vector amplification with low fidelity polymerases and the cost of synthesizing longer oligos, which...simple cloning procedures. Use high fidelity DNA polymerase This will minimize random errors during PCR: ...aliquots of PCR master-mix, containing buffer, polymerase and dNTPs, can be stored frozen: just defrost...mutagenesis by placing homologous ends on DNA using polymerase chain reaction." Biotechniques 10.1 (1991): 62...

  19. Getting the Most from Your Lentiviral Transduction

    Type
    Blog Post
    ...positively charged polycation, polybrene. Interestingly, one study found that alternative polycations such as DEAE-dextran...efficiency of lentiviral vectors: effect of sera and polycations.” Molecular Biotechnology 53.3 (2013). 308-14...

  20. The Fluorescent Vegetables in Aptamer Soup

    Type
    Blog Post
    ...developed to quantify RNA transcription by RNA Polymerase III. By expressing Corn under promoters for transcripts... transcripts uniquely transcribed by RNA Polymerase III (5S, tRNA and U6) and measuring Corn fluorescence...effect of mTOR inhibition on the activity of RNA Polymerase III. Red and orange fluorescent aptamers Corn...PMC5602116. Song, Wenjiao, et al. "Imaging RNA polymerase III transcription using a photostable RNA–fluorophore...
Showing: 41 - 60 of 289 results