Use of 'long' polymerase chain reaction and magnetic beads for extraction of chromosome-specific cDNAs.
The detection of expressed sequences of genomic DNA is an important aspect of the human genome project. A technique is described where 'long' polymerase chain reaction (LPCR), which allows for extended large fragment production of > 10-20 kb, is used with Alu primers to generate a biotinylated template for cDNA hybridization. Streptavidin-coated magnetic beads are used to extract the long PCR templates and bound cDNAs, which are recovered by standard PCR. This method allows the isolation of cDNAs from virtually any human DNA source and should be valuable in expression mapping, positional cloning and gene isolation.
Liang
B C
BC
Clinical Pharmacology Branch, National Cancer Institute, National Institutes of Health, Bethesda, MD, USA.
D001483:Base Sequence; D002877:Chromosomes, Human; D018076:DNA, Complementary / Q000302:isolation & purification*; D015894:Genome, Human; D006801:Humans; D006822:Hybrid Cells; D017404:In Situ Hybridization, Fluorescence; D008280:Magnetics; D008863:Microspheres; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods*
D018076:DNA, Complementary