An improved PCR-based amplification of unknown homologous DNA sequences.
The PCR primers used for cloning of evolutionary conserved genes or homologous DNA sequences are usually guessmer oligonucleotides. We introduce a simple way using Pfu polymerase to overcome possible PCR amplification failure because of 3'-end mismatches of guessed primers with the target DNA.
Naghibalhossaini
Fakhraddin
F
Department of Biochemistry, Shiraz University of Medical Sciences, Zand Street, Shiraz 71345, Iran. fakhraddin.naghibalhossaini@elf.mcgill.ca
Moaddeb
Afsaneh
A
D000818:Animals; D015703:Antigens, CD / Q000235:genetics; D002272:Carcinoembryonic Antigen / Q000235:genetics; D015815:Cell Adhesion Molecules / Q000235:genetics; D003001:Cloning, Molecular; D004247:DNA / Q000737:chemistry* / Q000235:genetics; D017931:DNA Primers / Q000737:chemistry* / Q000235:genetics; D004259:DNA-Directed DNA Polymerase / Q000737:chemistry*; D006801:Humans; D016133:Polymerase Chain Reaction / Q000379:methods; D012689:Sequence Homology, Nucleic Acid
D015703:Antigens, CD; C069847:CD66 antigens; D002272:Carcinoembryonic Antigen; D015815:Cell Adhesion Molecules; D017931:DNA Primers; D004247:DNA; C072146:Pfu DNA polymerase; D004259:DNA-Directed DNA Polymerase