Subtraction of cap-trapped full-length cDNA libraries to select rare transcripts
The normalization and subtraction of highly expressed cDNAs from relatively large tissues before cloning dramatically enhanced the gene discovery by sequencing for the mouse full-length cDNA encyclopedia, but these methods have not been suitable for limited RNA materials. To normalize and subtract f...
| Publié dans: | BioTechniques. - 1991. - 35(2003), 3 vom: 30. Sept., Seite 510-6, 518 |
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| Auteur principal: | |
| Autres auteurs: | , , , , , , , , |
| Format: | Article |
| Langue: | English |
| Publié: |
2003
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| Accès à la collection: | BioTechniques |
| Sujets: | Comparative Study Evaluation Study Research Support, Non-U.S. Gov't Technical Report Validation Study Journal Article |
| Résumé: | The normalization and subtraction of highly expressed cDNAs from relatively large tissues before cloning dramatically enhanced the gene discovery by sequencing for the mouse full-length 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 lambda 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 |
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| Description: | Date Completed 11.05.2004 Date Revised 10.12.2019 published: Print Citation Status MEDLINE |
| ISSN: | 1940-9818 |