Method for Preparation of cRNA
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United States of America Patent
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Issued Date -
Jan 28, 2010
app pub date -
Dec 5, 2006
filing date -
Dec 5, 2005
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Abandoned
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Abstract
It is an object of the present invention to provide a method for preparing a cRNA, the method being capable of preventing decrease in cRNA yield. In order to achieve the object, the method comprises (a) a step of performing a reaction for preparing a single-stranded cDNA by treating with RNaseH an mRNA-cDNA hybrid prepared by reverse transcription and a reaction for preparing a double-stranded cDNA from the single-stranded cDNA and then inactivating the RNaseH contained in the resultant reaction solution; (b) a step of contacting the reaction solution with a solid support having a cationic group on its surface under pH conditions where the cationic group is positively charged; (c) a step of separating the solid support from the reaction solution; (d) a step of eluting the double-stranded cDNA from the solid support; and (e) a step of performing a transcription reaction for preparing a cRNA from the double-stranded cDNA.
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