An improved method for constructing a full-length enriched cDNA library using small amounts of total RNA as a starting material
Experimental & Molecular Medicine
;
: 586-590, 2003.
Article
in English
| WPRIM
| ID: wpr-84207
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 microg of total RNA. These libraries contained 8x10(5) to 8x10(6) 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.
Full text:
Available
Index:
WPRIM (Western Pacific)
Main subject:
RNA
/
Base Sequence
/
Gene Library
/
Cloning, Molecular
/
Molecular Weight
Language:
English
Journal:
Experimental & Molecular Medicine
Year:
2003
Type:
Article
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