Expression profiling of glycosyltransferases using RT-PCR.
The quantitative reverse-transcription polymerase chain reaction (RT-PCR) is a method used to quantify messenger RNA (mRNA) levels. It is particularly applicable to quantification of mRNAs that are transcribed at very low levels, such as glycosyltransferases (GTs). In this chapter, I describe preliminary experiments for obtaining conditions for a quantitative RT-PCR method to quantify transcript levels of GTs and related genes potentially involved in L-selectin-mediated lymphocyte homing in the mouse gastric mucosa infected by Helicobacter felis. This method was developed by modifying conventional RT-PCR protocols and does not require fluorescence-detecting thermal cyclers. The method described here is particularly useful for assaying large numbers of samples that require very accurate analysis.
Kobayashi
Motohiro
M
Department of Pathology, Shinshu University School of Medicine, Matsumoto, Japan.
D000818:Animals; D020869:Gene Expression Profiling / Q000379:methods*; D016695:Glycosyltransferases / Q000096:biosynthesis* / Q000235:genetics*; D051379:Mice; D012333:RNA, Messenger / Q000032:analysis / Q000096:biosynthesis; D015203:Reproducibility of Results; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000191:economics / Q000379:methods*
D012333:RNA, Messenger; D016695:Glycosyltransferases