...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 ...

...the delivery of genetic information to hard-to-transfect mammalian cells. In addition, "viral pseudotyping...

... Vector Core (Link opens in a new window) . Transfections are performed using the transfer plasmid, a ...

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

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

...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....

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

...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...

...restriction digest! Then, you can transform or transfect the cells that you want to use for your experiments...

...structures within cells. HeLa cells were transfected with plasmids encoding FPs tagged with the plasma...

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

...protein of interest, such as a sample transiently transfected or stably infected with the protein of interest...

...scientists use a multitude of techniques including co-transfection of two or more plasmids, the use of multiple...

...Virus Name Description Link to Video General Transfection Introduce plasmid DNA to mammalian cells Lentivirus...Produce lentivirus with a polyethyenimine (PEI) transfection protocol Fluorescence Titering Assay for Lentivirus...preparation Antibodies Name Description Link to Video Transfection Introduce plasmid DNA to mammalian cells to ...

... protein expression observed using transient transfection approaches, generating cell lines using lentiviral... stable cell line, as opposed to transiently-transfected cells, increases reproducibility, as it eliminates...variation associated with repeated transient transfection. Some lentiviral vectors deliver mammalian antibiotic...Different brands and FBS lots can promote or inhibit transfection. Test a variety of brands and FBS lot numbers...

...optimizing this step, or consider enriching for transfected cells by flow sorting or drug selection, assuming...

...plasmid, and (3) the packaging plasmid. These are transfected into target cells which produce lentiviral particles...

... as early as 48-72 hours post-transformation/transfection in a transient assay. For stable transformation...