...percentage of fluorescent cells, these cells can still be used in downstream applications. Safety Warnings...ensure that you are in compliance with your institution’s biosafety regulations. Workflow Timeline Day...with multiple integration events leading to underestimation of the true titer. Calculate the transduction...

...genome-containing virions. Phenol red is added to clearly distinguish the steps. This method will help enrich the prep...ultracentrifugation. The steps are clearly distinguishable thanks to the use of phenol red. Image adapted...cartoon indicating the position of the needle for harvesting of the purified AAV using option #1. Right image...

... bath Pipetman Reagents 37 g pre-mixed powder consisting of: 5.0 g yeast extract 10.0 g peptone from casein...mL of agar and on which you can individually distinguish a maximum of ~100 colonies Pro-Tip If you will...After overnight drying, we place the plates in a plastic bag with an absorbent material to reduce condensation...

...lentiCas9-Blast 1 and then selected with 1 μg/mL blasticidin for 9 days. Single cells were then plated in ...transduced (MOI = 37) and selected with 1 μg/mL blasticidin for 9 days. Single cells from a stable cell pool...then expanded for an additional 3 weeks under blasticidin selection. Anti-Cas9 Western blotting was performed...

...ensure that you are in compliance with your institution’s biosafety regulations. Last Update: July 7, ... µg/mL. Mix well and incubate 72 h. Benzonase Digestion and Cell Harvest Gently aspirate media from the... When analyzing data there should be a clear distinction between negative droplets (black) and positive...

...for your eyes!), remove it from any gel tray as plastic will block much of the UV and with a clean, sterile... gel on a glass plate if available. Unlike the plastic tray, this will not significantly reduce the UV...

...Plasmid You may also like... How to Design a Primer Diagnostic Restriction Digest Introduction Sequence verification...the peaks in the area and make sure they are justifiable peaks. For instance, in the trace file below...

...recognition sequence. This can create multiple distinct overhangs with a single enzyme, and remove the...now look like this: Note: Elute your purified digestion and PCR products with sterile water (instead of...

...been placed in the centrifuge before you and may still be on the equipment. Before using the centrifuge...centrifuge. Immediately stop the centrifuge and investigate the issue. When you are finished spinning your...

...BSL-2). The purpose of the four levels is to distinguish between different types of hazards and to provide...with additional precautions to prevent injuries, ingestion, and exposures to hazardous materials. Video Watch...

...titers were similar to those reported by these institutions. This protocol is for a 96-well plate with 20...reaction volume. Last Update: February 13, 2019 Estimate of time required: ~3 h Workflow Timeline Plate...

...should be properly disposed of following your institution’s hazardous waste procedures. Last Update: February...desired concentration. If the sample concentration is still too low, repeat the centrifugation until the volume...

...ensure that you are in compliance with your institution’s biosafety regulations. Equipment Class II, Type... When analyzing data there should be a clear distinction between negative droplets (black) and positive...

...entire tube is dried but while the white pellet is still visible. Resuspend RNA pellet in RNase-free water...

... the absorbance of a liquid you can accurately estimate the concentration of a substance in that liquid...

...spreading of antibiotic makes it easy for an investigator to conveniently plate and select transformed...

...the scale later on as otherwise, the paper could stick to the surface. Tare the weighing boat or paper....

...of powder into 100 mL of deionized water. While stirring, slowly add hydrochloric acid until the solution...

...time by shortening or skipping many steps and will still get enough colonies for your next step. Remember...

...Remove the paraformaldehyde and follow your institution's laboratory safety guidelines for disposing of...