...data there should be a clear distinction between negative droplets (black) and positive droplets (blue/green...Lentivirus sample data, RRE positive (blue) and negative (black) droplets Figure 2: ddPCR Lentivirus sample...sample data, RPP30 positive (green) and negative (black) droplets Sample ID Plasmid ID Dilution Copies...

...replication competent virus, you should not serially propagate your virus as the chance of recombination events...

... Culture Purifying DNA from an Agarose Gel DNA Ligation Introduction Restriction enzyme digestion takes... fill in the overhangs and conduct a blunt end ligation. Use T4 DNA Polymerase or Klenow DNA Polymerase...

...DNA, the plasmid should also be propagated in an endonuclease negative E. coli strain such as NEB stable...

...DNA, the plasmids should also be propagated in an endonuclease negative E. coli strain such as NEB stable...

...2 weeks. All of the cells in the untransduced (negative) control well should be killed and no colonies...appropriate antibiotic. When the untransduced (negative control) cells have died and colonies are visible...

...shaking platform. Dilute the horseradish peroxidase-conjugated secondary antibody to the desired concentration...transfected with the protein of interest, and a negative control sample that does not express the protein...

...CGGCCTCAGTGAGCGA ITR Reverse Primer: 5’-GGAACCCCTAGTGATGGAGTT ITR Probe: -FAM-CACTCCCTCTCTGCGCGCTCG-BBQ...data there should be a clear distinction between negative droplets (black) and positive droplets (blue)....

...carefully bring a centrifuge into a cold room for centrifugation. Once you’re done using the centrifuge, bring...

...corresponds completely to the template DNA strand so elongation can proceed. Usually a guanine or cytosine is...

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

...of Plasmid DNA Agarose Gel Electrophoresis DNA Ligation Background Information Gel purification allows...

...reagents, determine the amount that you need to weigh. Gather the reagent you will be weighing out, a weighing...

...way to control which orientation the insert is ligated into the vector backbone and therefore the only...

... the density of your culture if needed. A good negative control is LB media + antibiotic without any bacteria...

...cells AAV Purification by Iodixanol Gradient Ultracentrifugation Protocol Mulitchannel Pipetting Technique...

... visible in the room and on the door. BSL-2 centrifugation steps require an aerosol-tight lid and rotors...

...enzymes, the product of a Gibson Assembly is a fully ligated double-stranded DNA molecule. This has proven to...

...plasmid to express the protein of interest, and a negative control sample such as cells that do not express...

... changes to the amount you are dispensing can negatively impact your experimental results. This protocol...