Genomic sequencing by ligation-mediated PCR.
Genomic sequencing permits studies of in vivo DNA methylation and protein-DNA interactions, but its use has been limited due to the complexity of the mammalian genome. Ligation-mediated PCR (LMPCR) is a sensitive genomic sequencing procedure that generates high quality, reproducible sequence ladders starting with only 1 microgram of uncloned mammalian DNA per reaction. This genomic sequencing procedure can be adapted for various methylation, in vivo footprinting and DNA adduct mapping procedures. We provide a detailed protocol for genomic sequencing by LMPCR and discuss the principles and applications of the method.
Pfeifer
G P
GP
Department of Biology, Beckman Research Institute of the City of Hope, Duarte, CA 91010, USA.
Riggs
A D
AD
D018736:DNA Adducts; D018983:DNA Footprinting; D019175:DNA Methylation; D004591:Electrophoresis, Polyacrylamide Gel; D009693:Nucleic Acid Hybridization; D016133:Polymerase Chain Reaction / Q000379:methods*; D017422:Sequence Analysis, DNA
D018736:DNA Adducts