Semi-Multiplex PCR Technique for Screening of Abundant Transcripts During Systematic Sequencing of cDNA Libraries
| Autor: | B. Pacchioni, S. Trevisan, S. Gomirato, S. Toppo, G. Valle, G. Lanfranchi |
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| Jazyk: | angličtina |
| Rok vydání: | 1996 |
| Předmět: | |
| Zdroj: | BioTechniques, Vol 21, Iss 4, Pp 644-649 (1996) |
| Druh dokumentu: | article |
| ISSN: | 1940-9818 0736-6205 |
| DOI: | 10.2144/96214st01 |
| Popis: | The systematic sequencing of cDNA libraries is an efficient approach for the identification of new genes, but the presence of abundant mRNAs is often a major problem. This paper describes a very simple method of “semi-multiplex PCR” that allows specific identification of such abundant transcripts before DNA sequencing without using nonrepresentative subtracted libraries. The PCR utilizes a series of forward primers specific for abundant transcripts with a pair of universal primers used for template generation. cDNA clones corresponding to abundant mRNAs are then revealed by double bands in agarose gel. |
| Databáze: | Directory of Open Access Journals |
| Externí odkaz: |
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