We narrowed to 960 results for: Cre

...transforming into bacteria D. Screening for Inserts D.1 Recommended materials D.2 Screening for inserts E. Producing...greater yield of DNA. Back to Top D. Screening for Inserts You may screen for plasmids that were successfully...prevailing guidelines when the pLKO.1 vector was created. Additional design rules can be found at the Broad...prevailing guidelines when the pLKO.1 vector was created. For alternative options, please refer to the enzyme...long-wavelength UV light. Short wavelength UV light will increase the chance of damaging the DNA. Purify the DNA...R0101S NcoI NEB #R0193S Agarose Sigma #A9539 D.2 Screening for Inserts Day 1: 1. Inoculate 5 colonies from...that gives you the optimal viral production. d. Create a master mix of FuGENE® 6 transfection reagent ...

...each (these can be built into PCR primers when creating a new part, with an additional 5' clamping sequence...Features: Modified RFC 10 w/ start and stop codons; Creates in-frame fusions w/ linker Thr-Gly; Use N-parts...

...generate viral vectors for research and gene therapy. Created with BioRender.com. Wild-type viruses have one ...vehicles to deliver genetic cargo into cells. To create safe and effective viral vectors, wild-type viral...

...used to reconstitute self-replicating microbes or recreate the diseases they cause. The standard format for...system - Wittbrodt Lab MultiSite Gateway Kit : Creating three-part fusion proteins - Planas and Santalucia...

... used in the creation of the hiPS cell lines The following plasmids can be used to create cell lines with...

...pooled libraries are useful tools for genetic screening experiments. These libraries target the solute...window) PMID: 40355755 (Link opens in a new window) Credits Contributing Authors Written by researchers in ...

...Pooled CRISPR Libraries for Screening Use pooled CRISPR libraries to screen for genes involved in specific...

...imaging microscopy (FLIM) or FLIM-FRET, where the increased energy transfer through FRET reduces the donor...by Eric Greenwald and the Jin Zhang Lab at UCSD Credits Contributing Authors Written and reviewed by the...

...help to increase the density of the culture. Normally cultures shake at 150–250 rpm, increase this to ...Autoclave and allow to cool to room temperature. Now screw on the top of the bottle and store the LB at room... storage of the bacteria, you can proceed with creating a glycerol stock . You can now isolate your plasmid...and with growth (bottom). Preparing Antibiotics Create a stock solution of your antibiotic. Unless otherwise... 2: High versus low copy plasmids in bacteria. Created with BioRender.com. I didn't get any growth after...Streaking and Isolating Bacteria on an LB Agar Plate Creating Bacterial Glycerol Stocks for Long-term Storage...

...about what DNA segments you will need to join to create your final plasmid. Adjacent segments should have...the figures). These identical sequences can be created via PCR with primers that contain a 5′ end that...split an antibiotic resistance gene to effectively create an extra part, one part has half of the antibiotic...unique function for the reaction: T5 Exonuclease - creates single-strand DNA 3’ overhangs by chewing back ...complementary DNA fragments, removing any nicks and creating a contiguous DNA fragment. Pro-Tip Add Extreme...make your own. Transform bacteria with the DNA and screen for the correct plasmid product by restriction ...reaction. However, some labs have observed a sharp decrease in success rate when assembling more than five...

...differences create contrast , which allows you to distinguish objects within the sample. You can increase contrast...resolution (the ability to distinguish between two discrete objects). Figure 1 depicts an image of a compound...Diagram of a compound light microscope with labels. Created with BioRender.com. Base Light Source Condenser...objective is in position over your sample. As you increase magnification, you will notice that the objectives...will likely need to re-adjust lighting and focus. Increase the power of the light source by opening the diaphragm...to bring the sample into focus. Pro-Tip As you increase your magnification, remember that the objectives...

...at the central position. Introducing Hi-P for screening phosphorylation-dependent protein-protein interactions...domain of NEDD4-2 (Pooled Libraries #111705-8) . Screening kinases The Mode #1 phosphosite library can also...

...polymerase/exonuclease functions, T4 DNA polymerase can create overhangs of varying length (typically 10-12 bp...distance from their recognition sequence. This can create multiple distinct overhangs with a single enzyme...concentrations in subsequent reactions. Step 3: Create Vector Overhangs Treat the linearized vector with...may be accomplished by gel purification . Step 5: Create Insert Overhangs Treat your purified PCR product...first base removed by the restriction digest to create the free 5' end of the cut vector (shown in blue...across multiple concentrations of insert, thereby increasing your chances of success. Set up a vector only...

...available directly from the labs in which they were created. Find their contact information by following the...

...single copy of DNA (the template), a researcher can create thousands of identical copies using a simple set...oligo primers. This molecular biology technique creates several micrograms of target DNA from just a few...: If you are doing multiple PCRs, save time by creating a “master mix,” which minimizes the number of ...FAQ Tips If your target DNA sequence is GC-rich, increase the time of the denaturing. Your 5’ and 3’ primers... If you are getting non-specific PCR products, increase the annealing temperature step-wise by 1-2°C. ...DNA mutagenesis. A higher cation concentration increases the error rate of the DNA polymerase. Deoxynucleoside...

...collection of Addgene's video content, including how-to screen captures, lab procedure protocols, and career advice... Description Related Page Making LB Agar Plates Create plates to culture bacteria in the lab Pouring LB...with CRISPR Protocol Over-Agar Antibiotic Plating Create plates to culture bacteria in the lab Over-Agar...tips and tricks to reduce transformation time and increase efficiency Bacterial Transformation Protocol Inoculating... from an Agarose Gel Protocol How-To Videos Screencapture walk-throughs to help guide you through Addgene's...

...dramatically increases the chances of individual plasmids being lost from the pool and/or increasing the skewness...plasmids in a single sample. They are often used for screening, barcoding, or other high throughput multiplexed...pipettes Ice slurry (Ice bucket with ice and water to create slurry) 100 mL LB At Addgene we use premixed LB...Immediately purify them! Commercial Maxipreps rely on incremental, ordered cell lysis. E. coli cells are subject...spreading less bacteria on a given plate (example: by increasing the total number of bioassay plates used). Scaling...

...well in sparse/individual cultures due to lack of secreted growth factors, so not every cell type will be...cell growth, such as using culture medium with increased serum concentrations, may be optimal for different...monoclonal lines have been sufficiently expanded, screen the lines for transgene expression and/or other...phenotypes. For example, perform Western blotting to screen for lines with the highest or lowest transgene ... vectors and genome-wide libraries for CRISPR screening. Sanjana NE, Shalem O, Zhang F. Nature Methods...

...concentration of positive droplets decreases by a factor of ~2. To increase the accuracy of the titer, take...the 96-well plate sealer by gently touching the screen. Prepare the Serial Dilution Place a 48-well dilution...the plate sealer is not at temperature, touch the screen on the plate sealer to allow it to get to temperature...therefore the concentration obtained by ddPCR should decrease by a factor of 2 across the dilutions. In the ...

...the 96-well plate sealer by gently touching the screen. Preparation of the Master Mix Place a ddPCR plate...the plate sealer is not at temperature, touch the screen on the plate sealer to allow it to get to temperature... concentration of RRE positive droplets should decrease by a factor of 2 across the dilutions. RPP30 copies...below, the concentration of RRE positive droplets increases by a factor of ~2 as you progress from the higher...stays relatively even across samples (green). To increase the accuracy of the titer, calculate an average...