...Lentiviral Particles E.1 Recommended materials E.2 Protocol for producing lentiviral particles F. Infecting...for producing viral particles. pMD2.G Envelope plasmid for producing viral particles. Note: pLKO.1 can ...selection G. Safety H. References H.1 Published articles H.2 Web resources I. Appendix I.1 Sequence of ...transfection, or can be converted into lentiviral particles for subsequent infection of a target cell line...sequence. Back to Top E. Producing Lentiviral Particles Before this step, you must contact your institution...Addgene recommends that you generate lentiviral particles and infect the target cells. Addition of puromycin...Tools page. E.2 Protocol for Producing Lentiviral Particles This protocol is for transfection in a 6 cm plate...

...B): LiCl selectively precipitates RNA versus DNA or proteins. Add the correct amount of 7.5 M LiCl solution...solution to bring the concentration of LiCl in the extracted aqueous layer to 2.5 M. Incubate at -20°C ...

...for the presence of particles still in solution. Continue to stir until all particles have dissolved. This...stir on a magnetic stir plate. Stir until all particles have dissolved. Note: The powder should dissolve...

...containing) particles. On average, iodixanol-purified AAV preparations contain ~20% empty particles. 60% iodixanol...

...proteins, cell fragments, salt and other extra particles from solutions. Supernatant contains the plasmid...may appear, consisting of precipitated protein particles Bottom phase - Organic phase (protein) Pipet the...

...too many cells relative to the number of virus particles reduces the transduction efficiency, resulting... so many that they vastly outnumber the virus particles. Incubate the cells with the virus for 48–72 h...

...Determine the number of genome-containing particles in an AAV prep using qPCR, SYBR Green Technology,... to determine the number of genome-containing particles of an AAV prep using SYBR green technology. This...

...generally represents the number of infectious viral particles. Users may need to run lower or higher dilutions... zero (A01). In this protocol, the lentiviral particles are serially diluted and used to transduce HEK293T...

...generally represents the physical number of viral particles. Users may need to run lower or higher dilutions...

...filters made of materials other than PES. AAV particles stick to many other surfaces, but do not stick...

... are not efficiently packaged into lentiviral particles. See below for options if the recombinant band...