Improved PCR method for amplification of GC-rich DNA sequences.
Most housekeeping genes, tumor-suppressor genes, and approx 40% of tissue-specific genes contain G+C sequences in their promoter region that were very difficult to amplify. In this report, we propose an improved polymerase chain reaction (PCR) method to be used for successful amplification of the tissue factor pathway inhibitor (TFPI)-2 gene promoter region that exhibit >70% G+C content in a sequence of approx 300 bp and a complete CpG island region spanning exon 1, the three transcription initiation sites, and the translation start site. Therefore, this method can be recommended to amplify other GC-rich genomic templates.
Hubé
F
F
Inserm, U618, Tours, F-37032 France; IFR 135, Tours, F-37044 France.
Reverdiau
P
P
Iochmann
S
S
Gruel
Y
Y
D001483:Base Sequence; D018899:CpG Islands; D004256:DNA Polymerase I; D004121:Dimethyl Sulfoxide; D005559:Formamides; D020862:GC Rich Sequence; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods*; D011401:Promoter Regions, Genetic / Q000235:genetics*; D017422:Sequence Analysis, DNA; D012997:Solvents
D005559:Formamides; D012997:Solvents; C031066:formamide; D004256:DNA Polymerase I; D004121:Dimethyl Sulfoxide