...cells in CS2 Day 2: Seed cells in CS5 Day 3 (am): Transfect cells Day 7 (am): Harvest cells Equipment Class...brands and lots of FBS can promote or inhibit transfection. Test a variety of brands and lots of FBS to...Polyethylenimine (PEI) 25 kDa MW Pro-Tip Other transfection reagents may be used in this protocol, but their...their conditions must be optimized. Plasmids for transfection: pHelper; pRC (Rep-Cap), plasmid expressing ... Pass and plate the cells the day before the transfection. Thaw a new vial of cells after 30 passages....to the incubator for 24 h–36 h. Proceed with transfection: Calculate the amount of each plasmid needed...Note that longer incubation times can reduce transfection efficiency. Add the OptiMEM + DNA + PEI solution...

...Transformation, Transduction, Bacterial Conjugation, and Transfection Resources on Addgene.org Addgene's Synthetic...

...over for other applications, like PCR, cloning, transfection, or long-term storage. But what about those ...

...Day 2: b. Perform the transfection in the late afternoon because the transfection mix should only be incubated...pLKO.1 can be introduced into cells via direct transfection, or can be converted into lentiviral particles...transient knockdown of protein expression, you may transfect plasmid DNA directly into the target cells. The...#12259 HEK-293T cells GenHunter: #Q401 FuGENE® 6 Transfection Reagent Roche Applied Biosciences: #11814443001...Producing Lentiviral Particles This protocol is for transfection in a 6 cm plate. The protocol can be scaled ... as needed. Day 1: a. For each plasmid to be transfected, plate 7×105 HEK-293T cells in 5 mL of media ...polystyrene tubes), make a cocktail for each transfection: 1 μg pLKO.1 shRNA plasmid 750 ng psPAX2 packaging...

...electroporation or poor cellular entry with other transfection methods. The Junwei Shi Lab addressed this problem...

...loci within a single cell. One might imagine transfecting cells with both the nuclease-positive and nuclease-null...

...tag. The two PCR reactions are then mixed and transfected along with a Cas9 expression vector. They can...

...too young and should not be used for important transfection experiments right after they’ve been thawed....

... adenoviral stock (rAdV-S). HEK293 cells are transfected with the AdV plasmid construct from step #1 and...

...could interfere with interpreting the results of transfection assays. Go forth and perform your DNA extractions...