[High performance liquid chromatography for analysis of reverse transcription polymerase chain reaction (RT-PCR) products].
The polymerase chain reaction is a powerful method for amplifying specific DNA sequences in vitro. Reverse transcribing mRNA into cDNA expands the use of PCR to monitor mRNA expression in biological system. A method for the analysis of RT-PCR products by HPLC was developed. The separation was performed on a nonporous ion exchange resin column with gradient elution of sodium chloride in 20 mmol/L Tris-HCl buffer (pH 9.0) at a flow rate of 1.0 mL/min and the detection wavelength was 260 nm. lambda-DNA-Hind III digest and a series of RT-PCR products were analyzed for studying the mRNA expression of secreted phospholipase A2 after being injured.
Liao
J
J
Medical Experiment & Analysis Center of PLA General Hospital, Beijing 100853, China.
Zhao
Y
Y
Dong
F
F
Yang
J
J
Hao
X
X
D000818:Animals; D002851:Chromatography, High Pressure Liquid; D003250:Constriction; D007422:Intestines / Q000098:blood supply* / Q000201:enzymology; D008297:Male; D017537:Mesenteric Artery, Inferior; D010741:Phospholipases A / Q000096:biosynthesis* / Q000235:genetics; D054467:Phospholipases A2; D012333:RNA, Messenger / Q000096:biosynthesis / Q000235:genetics; D051381:Rats; D017208:Rats, Wistar; D015427:Reperfusion Injury / Q000201:enzymology*; D020133:Reverse Transcriptase Polymerase Chain Reaction
D012333:RNA, Messenger; D010741:Phospholipases A; D054467:Phospholipases A2