Application of quantitative real-time PCR for monitoring the process of enrichment of clones on in vitro protein selection.
In vitro selection of proteins from cDNA libraries using display technologies, such as the in vitro virus method, is a powerful means for the discovery of novel protein interactions. After iterative screening, selected proteins are usually identified and evaluated by cloning and sequencing analysis. Previously we applied real-time PCR for evaluation of the sequences obtained on in vitro virus screening. Here, we have presented additional data regarding monitoring of the process of enrichment of selected clones in each round of selection and elimination of false positives by real-time PCR, and have also discussed the utility of the novel method. This approach should also be applicable to other display technologies.
Horisawa
Kenichi
K
Department of Biosciences and Informatics, Faculty of Science and Technology, Keio University, 3-14-1 Hiyoshi, Kohoku-ku, Yokohama 223-8522.
Doi
Nobuhide
N
Takashima
Hideaki
H
Yanagawa
Hiroshi
H
D018076:DNA, Complementary / Q000032:analysis*; D015723:Gene Library; D016133:Polymerase Chain Reaction; D011506:Proteins / Q000235:genetics*; D017422:Sequence Analysis, DNA / Q000379:methods
D018076:DNA, Complementary; D011506:Proteins