Codons corresponding to asparagine residues of RTAs were mutated to aspartate (D) or glutamine (Q) using the QuikChange Site-Directed Mutagenesis Kit (Stratagene) and confirmed by sequencing. For transfection with 293T cells, calcium phosphate or polyethylenimine was used when cells were of ~50 to 60% confluency. For BAC DNA transfection in SLK cells, Lipofectamine 3000 was used according to the manufacturer's instructions. Cells in each well of a 24-well plate were transfected with approximately 1 μg of BAC DNA, and SLK/iBAC-positive cells were selected with hygromycin B (200 μg/ml) at 48 hours after transfection.

Note: The content above has been extracted from a research article, so it may not display correctly.

Please log in to submit your questions online.
Your question will be posted on the Bio-101 website. We will send your questions to the authors of this protocol and Bio-protocol community members who are experienced with this method. you will be informed using the email address associated with your Bio-protocol account.

We use cookies on this site to enhance your user experience. By using our website, you are agreeing to allow the storage of cookies on your computer.