Interference by complex structures of target DNA with specific PCR amplification.
It has been considered that target DNA is a forgiving component for PCR amplification. Herein we present evidence to demonstrate that secondary structure located at the end of a template may interfere with the specificity of amplification. Experiments indicate that nonspecific amplification results from a long stretch of stem and loop structures at the 3' end of prochymosin cDNA. Based on the sequence of mRNA coding for prochymosin, it is argued that the sequence responsible for the formation of the complex structure described here is most likely generated during synthesis of the second cDNA strand.
Zhang
Z
Z
Chinese Academy of Sciences, Institute of Biophysics, Beijing, China.
Huang
K
K
Zhang
Y
Y
Liu
N
N
Yang
K
K
D001483:Base Sequence; D012085:Chymosin / Q000235:genetics*; D018076:DNA, Complementary / Q000737:chemistry*; D004591:Electrophoresis, Polyacrylamide Gel; D004792:Enzyme Precursors / Q000235:genetics*; D008969:Molecular Sequence Data; D009690:Nucleic Acid Conformation; D016133:Polymerase Chain Reaction; D013698:Templates, Genetic
D018076:DNA, Complementary; D004792:Enzyme Precursors; C032443:prorennin; D012085:Chymosin