Prevention of cross-contamination during sampling procedure in molecular detection for cancer micrometastasis.
Reverse transcriptase-polymerase chain reaction (RT-PCR) techniques have been widely employed as an ultra-sensitive method for detection of micrometastases in patients with various types of malignancies. Messenger RNA of a specific marker gene is a target for RT-PCR amplification to examine the presence of micrometastases in body fluids or tissues obtained from human. We developed the RT-PCR assay specific for rat beta-actin mRNA, which cannot detect human counterpart and assessed how much contamination of rat tissues can influence the result of RT-PCR assay and how to avoid the influence of the contamination in RT-PCR assay.
Fujiwara
Y
Y
Department of Surgery and Clinical Oncology, Graduate School of Medicine, Osaka University, 2-2 Yamada-Oka, Suita, Osaka, Japan. fujiwara@surg2.med.osaka-u.ac.jp
Ooka
M
M
Sugita
Y
Y
Sakita
I
I
Tamaki
Y
Y
Monden
M
M
D000199:Actins / Q000032:analysis* / Q000235:genetics; D000818:Animals; D004866:Equipment Contamination; D006801:Humans; D009362:Neoplasm Metastasis; D011786:Quality Control; D012333:RNA, Messenger / Q000032:analysis; D051381:Rats; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*
D000199:Actins; D012333:RNA, Messenger