PCR-based method for isolation of full-length clones and splice variants from cDNA libraries.
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.
Alphey
L
L
University of Manchester, School of Biological Sciences, England, UK. lalphey@fs2.scg.man.ac.uk
D000818:Animals; D003001:Cloning, Molecular; D017931:DNA Primers; D018076:DNA, Complementary / Q000235:genetics / Q000302:isolation & purification*; D004330:Drosophila; D004591:Electrophoresis, Polyacrylamide Gel; D015723:Gene Library; D016547:Kinesins / Q000235:genetics; D016133:Polymerase Chain Reaction / Q000379:methods*; D012326:RNA Splicing
D017931:DNA Primers; D018076:DNA, Complementary; D016547:Kinesins