Application of FLUPD-DD-PCR to the study of mRNA expression of glioma cells cultured under the condition of serum starvation

Fluorescently labeled universal primer directed differential display polymerase chain reaction technique (FLUPD-DD-PCR), an improved DD-PCR, is reported for the study of mRNA expression of glioma cells cultured with the serum starvation. The fluorescently labeled universal primer (FLUP) facilitates analysis of PCR products on an auto-sequencer. Compared with the traditional DD-PCR, FLUPD-DD-PCR has advantages of separation of target bands and measurement of mRNA expression quantitatively by adopting Cy5-labeled universal primer. And then the silver staining method has been used to recover the bands for use as template in re-pcr. In this study, 4 differential ESTs, of which 3 are novel ESTs, have been obtained. This work indicates some special novel genes, which can be induced to express under serum absent condition, are involved in coping with serum starvation stress on glioma cells growth.