| Status | 已發表Published |
| Generation of Longer cDNA Fragments from SAGE Tags for Gene Identification | |
Wang, S. M.
| |
| 2002 | |
| Source Publication | Generation of cDNA Libraries |
| Publication Place | Totowa |
| Publisher | HUMANA PRESS Inc. Totowa, New Jersey |
| Pages | 207-222 |
| Abstract | SAGE (Serial Analysis of Gene Expression) is a powerful technique for genome-wide analysis of gene expression (1-14). However, almost two thirds of SAGE tags cannot be used directly for gene identification for two reasons. First, many of SAGE tags match to multiple known expressed sequences due to the short length of SAGE tag sequences (12-14). Second, many SAGE tags do not match any known expressed sequences because the sequences corresponding to these SAGE tags have not been identified yet (2-14). These problems substantially diminish the power of the SAGE technique. The GLGI (Generation of Longer cDNA fragments from SAGE tags for Gene Identification) technique was designed to solve these two problems (15). The basic principle of GLGI is to use the SAGE tag as the sense primer, and anchored oligo dT primers as the antisense primer to amplify the original 3'cDNA from which the SAGE tag was derived. The size of 3' cDNA will be hundred bases longer which is long enough for solving the two problems. In a typical SAGE project, hundreds or thousands of SAGE tags need to be further analyzed. To facilitate such large-scale performance, we developed the GLGI method into a high-throughput procedure (16). In this high-throughput GLGI procedure, 3' cDNAs starting from the last CATG are used as the templates for GLGI amplification, a SAGE tag sequence is used as the sense primer and an universal sequence located at the 3' end of all the cDNA templates generated from anchored oligo(dT) primers is used as antisense primer to amplify the original 3' cDNA template from which the SAGE tag was derived. The whole process is simple, rapid and low-cost, with highly efficiency (12-14, 16). Besides from identifying the correct genes for SAGE tags with multiple matches, GLGI can be used for large-scale identification of novel genes by converting novel SAGE tags into novel 3 cDNAs. This GLGI method provides a powerful tool for gene identification in the human and other eukaryotic genomes. |
| Keyword | mRNA cDNA polyA gene |
| Language | 英語English |
| ISBN | 1588290662 |
| The Source to Article | PB_Publication |
| PUB ID | 43514 |
| Document Type | Book chapter |
| Collection | DEPARTMENT OF PUBLIC HEALTH AND MEDICINAL ADMINISTRATION |
| Recommended Citation GB/T 7714 | Wang, S. M.. Generation of Longer cDNA Fragments from SAGE Tags for Gene Identification[M]. Generation of cDNA Libraries, Totowa:HUMANA PRESS Inc. Totowa, New Jersey, 2002, 207-222. |
| APA | Wang, S. M..(2002). Generation of Longer cDNA Fragments from SAGE Tags for Gene Identification. Generation of cDNA Libraries, 207-222. |
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