Subtraction of cap-trapped full-length cDNA libraries to select rare transcripts
| Autor: | Tomoko Hirozane-Kishikawa, Toshiyuki Shiraki, Kazunori Waki, Mari Nakamura, Takahiro Arakawa, Jun Kawai, Michela Fagiolini, Takao K. Hensch, Yoshihide Hayashizaki, Piero Carninci |
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| Jazyk: | angličtina |
| Rok vydání: | 2003 |
| Předmět: | |
| Zdroj: | BioTechniques, Vol 35, Iss 3, Pp 510-518 (2003) |
| Druh dokumentu: | article |
| ISSN: | 1940-9818 0736-6205 |
| DOI: | 10.2144/03353st04 |
| Popis: | The normalization and subtraction of highly expressed cDNAs from relatively large tissues before cloning dramatically enhanced the gene discovery by sequencing for the mouse fulllength cDNA encyclopedia, but these methods have not been suitable for limited RNA materials. To normalize and subtract full-length cDNA libraries derived from limited quantities of total RNA, here we report a method to subtract plasmid libraries excised from size-unbiased amplified λ phage cDNA libraries that avoids heavily biasing steps such as PCR and plasmid library amplification. The proportion of full-length cDNAs and the gene discovery rate are high, and library diversity can be validated by in silico randomization. |
| Databáze: | Directory of Open Access Journals |
| Externí odkaz: |
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