Intercept-PCR, an improvement for elevating performance to find a new member of a certain gene family.
We have established a method by which the performance of reverse transcriptase coupled polymerase chain reaction (RT-PCR) for seeking a new gene is improved. The actual procedure is quite easy: it is only to add several specific oligonucleotides into the reaction mixture of the usual RT-PCR. To verify the effectiveness of this method is also easy: it is only to detect the PCR products in the preliminary experiment. The finding in the present study provides valuable information for gene cloning tactics.
Matsuda
S
S
Nagoya University School of Medicine, Dep. Mol. Pathogen., Japan. smatsuda@med.nagoya-u.ac.jp
Ichigotani
Y
Y
Okuda
T
T
Miyazaki
K
K
Yamamoto
T
T
Nimura
Y
Y
Irimura
T
T
Nakatsugawa
S
S
Hamaguchi
M
M
D001483:Base Sequence; D002460:Cell Line; D017931:DNA Primers; D018076:DNA, Complementary; D006801:Humans; D005810:Multigene Family; D016133:Polymerase Chain Reaction / Q000379:methods*; D012333:RNA, Messenger / Q000235:genetics; D015203:Reproducibility of Results; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012680:Sensitivity and Specificity
D017931:DNA Primers; D018076:DNA, Complementary; D012333:RNA, Messenger