Serial analysis of gene expression: rapid RT-PCR analysis of unknown SAGE tags.
In a pilot study on SAGE on Reed-Sternberg cells we have sequenced 1055 tags representing 701 genes. Screening of the GenBank database resulted in the identification of a corresponding gene or EST for 490 of them. For 211 of the tags no homology could be detected. A major problem of the serial analysis of gene expression (SAGE) approach is how to further analyse the unknown tags. We have developed an RT-PCR-based method, rapid analysis of unknown SAGE tags (RAST-PCR), to analyse the expression of the corresponding genes. This approach can be used as a screening method to investigate whether or not the gene is differentially expressed between several cell types of interest.
van den Berg
A
A
Department of Pathology and Laboratory Medicine, University Hospital Groningen, PO Box 30.001, 9700 Groningen, The Netherlands.
van der Leij
J
J
Poppema
S
S
D002460:Cell Line; D020224:Expressed Sequence Tags; D015870:Gene Expression; D010865:Pilot Projects; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D017422:Sequence Analysis, DNA; D014407:Tumor Cells, Cultured