Quantitation of glutaminase mRNA in enterocytes using competitive RT-PCR.
The concentration of mRNA within the intestinal mucosa is usually measured by either Northern blot analysis or semi-quantitative reverse transcription polymerase chain reaction (RT-PCR). However, these methods are limited by a lack of valid internal controls, low sensitivity, and large differences in the concentration of the internal control and target gene. The authors present an alternative method using competitive RT-PCR to measure glutaminase mRNA in isolated enterocytes.
The University Department of Surgery, Royal Perth Hospital, Perth, Australia.
S E
SE
Kong
K A
KA
Heel
J C
JC
Hall
R D
RD
McCauley
D000818:Animals; D001483:Base Sequence; D002138:Calibration; D003106:Colon; D017931:DNA Primers; D005972:Glutaminase / Q000235:genetics*; D007082:Ileum; D007413:Intestinal Mucosa / Q000201:enzymology*; D012333:RNA, Messenger / Q000032:analysis*; D051381:Rats; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods; D012680:Sensitivity and Specificity
D017931:DNA Primers; D012333:RNA, Messenger; D005972:Glutaminase