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Title | An improved method for constructing a full-length enriched cDNA library using small amounts of total RNA as a starting material |
Authors | Jung Hwa Oh; Yong Sung Kim; Nam-Soon Kim |
Publisher | Korean Society of Medical Biochemistry |
Issue Date | 2003 |
Citation | Experimental and Molecular Medicine, vol. 35, no. 6, pp. 586-590 |
Keywords | cDNA library; full-length; oligo-capping; total RNA; adenine; complementary DNA; messenger RNA; RNA; data base; diagnosis, measurement and analysis |
Abstract | We have developed an improved method for constructing a full-length cDNA library using small quantity of material by modifying the original oligo-capping method. In our devised method, total RNAs are used in sequential oligo-capping steps directly without preliminary mRNA purification. Using this method, we constructed full-length cDNA libraries from 100 μg of total RNA. These libraries contained 8 × 105 to 8 × 106 independent clones with average insert sizes of 2.0 kb. Moreover, the number of full-length cDNAs containing the translation initiation codon ATG in the constructed libraries was estimated to 60-70%. In addition, 54% of the known cDNAs had a longer 5′ end than the corresponding genes in the public database. Our results show that the method can be effectively used to construct full-length enriched cDNA libraries, especially, if starting material is limited. |
ISSN | 1226-3613 |
Appears in Collections | |
Registered Date |
2017-04-19 |
| Files in This Item: | Size | Format | |
|---|---|---|---|
| 4853.pdf | 267Kb | Adobe PDF | 
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