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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 OhYong Sung KimNam-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 libraryfull-lengtholigo-cappingtotal RNAadeninecomplementary DNAmessenger RNARNAdata basediagnosis, 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

1. Journal Articles > Journal Articles

Registered Date

2017-04-19


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