Open AccessPCR-Based Method for Isolation of Full-Length Clones and Splice Variants from cDNA Libraries
Author(s) -
Luke Alphey
Publication year - 1997
Publication title -
biotechniques/biotechniques
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.617
H-Index - 131
eISSN - 1940-9818
pISSN - 0736-6205
DOI - 10.2144/97223st02
Subject(s) - complementary dna , splice , cdna library , biology , selection (genetic algorithm) , genetics , isolation (microbiology) , genomic library , microbiology and biotechnology , computational biology , polymerase chain reaction , gene , bioinformatics , base sequence , computer science , artificial intelligence
Most cDNA library screening procedures do not distinguish between full-length and incomplete clones and therefore may yield incomplete cDNA fragments. Thus, there is a widespread need for a method allowing the efficient selection of full-length clones. I present a rapid, PCR-based method that allows the simultaneous screening of > 10 6 cDNAs. The longest cDNA is identified in the first step so that incomplete clones may be eliminated from study at this stage to save time. The method also facilitates the identification and isolation of rare splice variants from a background of a more abundant variant.