Author: Liao, Pei-Yu; Choi, Yong Seok; Dinman, Jonathan D.; Lee, Kelvin H.
Title: The many paths to frameshifting: kinetic modelling and analysis of the effects of different elongation steps on programmed –1 ribosomal frameshifting Document date: 2010_9_7
ID: wwq0sd2r_21
Snippet: Test strains were grown in 100 ml LB medium containing 100 mg/ml ampicillin in 500 ml flasks at 250 rpm and 37 C. After 24 h, 200 OD 600 units of cells were collected by centrifugation at 4000g and 4 C for 20 min. Cells were lysed and purified by Ni-NTA under native conditions according to the manufacturer's protocol (Qiagen). Purified protein samples were resolved by SDS-PAGE (10% w/v polyacrylamide). Gel band excision and in-gel trypsin digesti.....
Document: Test strains were grown in 100 ml LB medium containing 100 mg/ml ampicillin in 500 ml flasks at 250 rpm and 37 C. After 24 h, 200 OD 600 units of cells were collected by centrifugation at 4000g and 4 C for 20 min. Cells were lysed and purified by Ni-NTA under native conditions according to the manufacturer's protocol (Qiagen). Purified protein samples were resolved by SDS-PAGE (10% w/v polyacrylamide). Gel band excision and in-gel trypsin digestion were performed using a previously described standard method (33) .
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