title: A primer design strategy for PCR amplification of GC-rich DNA sequences.
abstract: OBJECTIVE
  To establish a primer design method for amplification of GC-rich DNA sequences.
  
  
  METHODS
  A group of 15 pairs of primers with higher T(m) (>79.7°C) and lower level ΔT(m) (<1°C) were designed to amplify GC-rich sequences (66.0%-84.0%). The statistical analysis of primer parameters and GC content of PCR products was performed and compared with literatures. Other control experiments were conducted using shortened primers for GC-rich PCR amplifications in this study, and the statistical analysis of shortened primer parameters and GC content of PCR products was performed compared with primers not shortened. A group of 26 pairs of primers were designed to test the applicability of this primer designing strategy in amplifications of non-GC-rich sequences (35.2%-53.5%).
  
  
  RESULTS
  All the DNA sequences in this study were successfully amplified. Statistical analyses show that the T(m) and ΔT(m) were the main factors influencing amplifications.
  
  
  CONCLUSIONS
  This primer designing strategy offered a perfect tool for amplification of GC-rich sequences. It proves that the secondary structures cannot be formed at higher annealing temperature conditions (>65°C), and we can overcome this difficulty easily by designing primers and using higher annealing temperature.
authors.affiliation: Department of Cytobiology, Southern Medical University, Guangzhou 510515, PR China.
authors.forename: Li-Yan
authors.initials: LY
authors.lastname: Li
authors.forename: Qiang
authors.initials: Q
authors.lastname: Li
authors.forename: Yan-Hong
authors.initials: YH
authors.lastname: Yu
authors.forename: Mei
authors.initials: M
authors.lastname: Zhong
authors.forename: Lei
authors.initials: L
authors.lastname: Yang
authors.forename: Qing-Hong
authors.initials: QH
authors.lastname: Wu
authors.forename: Yu-Rong
authors.initials: YR
authors.lastname: Qiu
authors.forename: Shen-Qiu
authors.initials: SQ
authors.lastname: Luo
mesh_terms: D017931:DNA Primers / Q000235:genetics*; D004586:Electrophoresis; D005091:Exons / Q000235:genetics; D020862:GC Rich Sequence / Q000235:genetics*; D016133:Polymerase Chain Reaction / Q000379:methods*
chemical_list: D017931:DNA Primers