We narrowed to 622 results for: erf

...Resources Inducible and repressible promoters are powerful tools to control gene expression and study biological...

...your purified virus. Nonetheless, we recommend performing a buffer exchange before using the purified AAV... carefully puncture the QuickSeal tube at the interface of the 60% and 40% gradient (see Figure 2) with...Avoid the proteinaceous material near the 40–25% interface. Pro-Tip As soon as the top of the tube is punctured...solution will flow from the needle set at the 40–60% interface. Make sure that the microcentrifuge tubes are ... the QuickSeal tube slightly below the 60–40% interface with an 18 ga needle attached to a 10 mL syringe...collecting the proteinaceous material at the 40–25% interface. Repeat for each QuickSeal tube. (Optional) For...ultracentrifugation. The arrow indicates the 60–40% interface. The vertical black line indicates the location...

...Follow this protocol to perform amplification of CRISPR pooled plasmid libraries in Escherichia coli ...Estimated time 2-3 hours) Transformation should be performed at the end of the day to ensure that growth times...shaking period, pool and gently mix the four tubes. Perform sequential 1:100 dilutions of the cells (add 10...10 µL of the pool to 990 µL LB then perform a second and then third 1:100 dilution). Plate 100 µL of each... recombined fraction but care must be taken to perform adequate NGS based QC to ensure no change in representation...to the pre-amplified stock. A last option is to perform a gel extraction of either the original sample ...of the DNA, but please note that NGS should be performed to ensure representation is maintained. Maxipreps...

...standard polymerase protocol), causing the enzyme to perform exonuclease activity until the first "G" in the...sterile dH20 to 40 μl Step 4: Amplify Insert by PCR Perform PCR amplification of your insert following the ...your PCR product before proceeding, as they will interfere with the exonuclease activity of T4 Pol in the...

...phone-sized, foldable microscopes that aren't very powerful (but are cheap and accessible) to massive transmission... are multiple objective lenses. Each objective performs essentially the same task but has different magnifying...scratches, and dust can easily impair microscope performance. Be sure to clean up your work area and microscope...

... follow the kit's instructions. If you want to perform plasmid purification without using a kit, you can...ribonuclease that digests single-stranded RNA. Optional: Perform phenol-chloroform extraction - see protocol below... DNA purified without a kit it is advisable to perform a phenol/chloroform extraction of the supernatant...

...Watch the protocol video below to learn how to perform Polymerase Chain Reaction (PCR). Basic PCR Program...pairs. The initial denaturation step is commonly performed at 94–98°C. Denature 30 seconds at 94°C: Continued...reagent by the number of individual PCRs you wish to perform and add 10% extra to account for pipetting error...

... of a stable cell culture after transduction. Performing antibiotic selection on transduced cells enables...5 dilutions plus one 'no virus' control well). Perform a "reverse transduction" by seeding 50,000 cells...untransduced well (0 µL lentivirus, above) are dying. Perform regular media changes and monitor the growth of...

...monoclonal cell lines. When this protocol was performed at Addgene on the A549 cells described in our ...expression and/or other phenotypes. For example, perform Western blotting to screen for lines with the highest...blasticidin selection. Anti-Cas9 Western blotting was performed on whole-cell extracts from the initial stable...

...Protocol, tips, and FAQ for how to perform a restriction digest of plasmid DNA...more enzyme is advisable. Reactions are often performed with 0.2-0.5 µL of enzyme because it is difficult...

...fragments. This reaction, called ligation, is performed by the T4 DNA ligase enzyme. The DNA ligase catalyzes...digested vector backbone with a phosphatase before performing the ligation reaction (phosphatase removes the...

...Standard protocol for performing agarose gel electrophoresis, including tips to improve resolution and...Watch the protocol video below to learn how to perform gel electrophoresis. Equipment Casting tray Well...

...function by RNA interference (RNAi) RNA interference (RNAi) is a powerful tool for performing targeted knockdowns...the students performing these experiments have a good baseline for comparison. Performing WISH using these...are too few resources to teach students how to perform these experiments in the beginning. In most classrooms...observation and dissections. Students don’t usually perform more modern techniques used in genetics or molecular...addition, the Sánchez lab has made it easier to perform advanced developmental biology experiments in planarians...genes. Double-stranded RNA (dsRNA)-mediated RNA interference (RNAi) to knockdown expression of specific gene...both WISH and RNAi experiments, as well as how to perform worm amputations. They also created an online companion...

...than one another, a two-tailed test should be performed. This test will assess if the mean of an experimental...tests, referred to as post-hoc analysis, must be performed. These will be discussed later. One and two-way...within your ANOVA/ANCOVA analysis. If you have performed a MANOVA/MANCOVA and want this information, multiple...multiple ANOVAs/ANCOVASs must be performed first, and then post hoc tests can be run on these tests. ...can reduce the family-wise error rate by only performing the comparisons you really care about. Family-wise... positive occurring when many comparisons are performed. For example, if your alpha is set at .05, then...test will be incorrect. If you have 5 samples and perform every pairwise comparison (10 in total), your likelihood...

...optimization as each antibody performs differently and some antibodies impact the performance of other antibodies...beautiful results. Performing an immunofluorescence experiment The process to perform IF can be broadly...important considerations to make when planning and performing an IF experiment. What is immunofluorescence... different species and colors makes it easy to perform multiplexed IF. By using multiple primary antibodies...in IF experiments. Precisely which controls you perform will depend on your experimental context.  Choosing... to use, then the primary antibody will likely perform as expected in both.  How do you find validated...may be, it is always important to verify their performance in your hands and in your specific experiment...

...pooled libraries have allowed scientists to easily perform genome-wide screens to effectively and efficiently...human genome-wide CRISPRko (knock out), CRISPRi (interference), and CRISRPa (activation) libraries that do...that improved sgRNA activity and thus library performance. The Root and Doench lab’s optimized genome-wide...CRISPRi, and CRISPRa libraries have been shown to outperform other popular CRISPR libraries (see below). Let...colon cancer) cells and assessed the libraries performance via the depletion of a gold standard set of genes...developed sgRNA-level metrics to assess library performance in viability screens. Based on these metrics ...sgRNAs, Brunello even with only a single sgRNA outperformed another CRISPR library with 6 sgRNAS, highlighting...

...protocols and performing rigorous quality control (QC). The specific QC experiments performed varies for ...of Pennsylvania Vector Core. Transfections are performed using the transfer plasmid, a plasmid encoding...and stored at -80 °C. Titer Titering is either performed by Addgene or by the University of Pennsylvania...Pennsylvania Vector Core. In general, titering is performed by the facility that produced the viral vector lot....Pennsylvania Vector Core is (as of April 2016) also performed by ddPCR.  Quality Control Addgene ensures high.... To learn which specific QC experiments were performed on your lot, please contact us. Full sequencing...the Viral Genome Next-generation sequencing is performed on viral genomes isolated from the final AAV preparation...

...is, an antibody shown to perform well in one set of conditions may not perform as well in other conditions...approach is RNA interference (RNAi). The most common method is transfection of short interfering RNA (siRNA...assumed (Ayoubi et al., 2023). Additionally, their performance depends on the specific protocol used and the...For example, we have found that some antibodies perform well in a PFA-saponin fixation and permeabilization...population (Figure 1), further optimization can be performed to both maximize the separation and reduce any... is possible to design experiments where the performance of the antibody can be assessed through its ability...different conjugates. Summary Flow cytometry is a powerful tool to delineate cell populations and to study...

...your antibody performs as expected in one application does not mean that it will perform as expected in...doesn’t necessarily mean that you yourself have to perform extensive validation experiments for every antibody...data, you may need to roll up your sleeves and perform some extra validation experiments yourself.  Goal...researchers can use to show that their antibody performs as expected. One of the benefits of these pillars...levels tend to correspond to protein levels, so you perform immunohistochemistry with your antibody on these... the epitope recognized by your antibody. You perform immunocytochemistry on these mutant cells as well...caveat is that different antibodies may or may not perform well under the same conditions. Example: You want...

...homemade competent cells of 107 CFU/μg are perfectly fine, yet performing multiple modifications will require... pathway to allow any cloning procedure to be performed using a simple two-step, 2 hr protocol prior to...adopted by any laboratory. All procedures are performed using the same three step protocol:  Fragment...efficiency of the strain: when asking the cell to perform multiple recombination events, all linear fragments... design (Figure 2), IVA cloning can be used to perform any plasmid modification, from insertions, deletions... reverse primer. Mutagenesis - Mutagenesis is performed by designing primers astride the old codon, amplifying...primers can be used in combination to simultaneously perform modifications, simply by adding all the necessary...