Patent · US Expired

Method and system for the preparation of cDNA

US6521411B2 · kind B2 · utility

25Cited by
24References
34Claims
0Family size

Assignee

Inventors

Key dates

Filing dateMar 13, 2001
Grant dateFeb 18, 2003
Priority date
Expiry dateMar 13, 2021

Classification

  • Technology area (CPC C)Chemistry; Metallurgy
  • CPC primaryC12N15/1096
  • WIPO fieldBiotechnology
  • WIPO sectorChemistry

Abstract

In one aspect, the present invention concerns an improved method for the preparation of cDNA libraries. Preferred embodiments of the invention include methods and systems capable of generating cDNA libraries enriched for high molecular weight cDNA inserts. The method generally entails the following steps: (1) size-based separation of a plurality of mRNA molecules by Ion-Pairing Reversed-Phase Chromatography (IP-RPC), preferably using HPLC under denaturing conditions; and (2) collection of a fraction of the mRNA molecules that is enriched for mRNA molecules of a desired size range (in a preferred embodiment of the invention, larger-sized mRNA molecules are collected, e.g., mRNA of length greater than 10 kb). The collected fraction of mRNA molecules can be reverse transcribed to form a library of cDNA inserts enriched for inserts of a desired relative size range.

Source: USPTO / EPO open patent data. Objective bibliographic and citation counts.