Method for making full-length cDNA libraries
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United States of America Patent
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Jan 16, 2001
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app pub date -
Nov 20, 1996
filing date -
Mar 18, 1996
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Abstract
Disclosed is a method for making full-length CDNA libraries, which is for making libraries of cDNAs having a length corresponding to a full-length of mRNAs and comprises the following steps of; binding a tag molecule to a diol structure present in 5' Cap (.sup.7Me G.sub.ppp N) sites of mRNAs, forming RNA-DNA hybrids by reverse transcription using primers such as oligo dT and the mRNAs connected with the tag molecule as templates, and separating RNA-DNA hybrids carrying a DNA corresponding to a full-length of mRNAs from the RNA-DNA hybrids formed above by using function of the tag molecule. To obtain mRNA connected with a tag molecule, the diol structure present in 5' Cap site of mRNA is subjected to a ring-open reaction by oxidation with sodium periodate to form a dialdehyde and the dialdehyde is reacted with a tag molecule having a hydrazine terminus. According to the present invention, there are provided a novel method capable of efficiently labeling 5' Cap site and a method for making full-length cDNA libraries utilizing the labeling method.
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