Patent ID: 6387624
Filing Date: 2002-05-14
Classification: C12N

Abstract:
A method of producing uni-directionally cloned complementary DNA libraries from mRNA, the method comprising:contacting mRNA with a primer mixture comprising a plurality of primers having different sequences, wherein each primer in the mixture comprises: at least 10 contiguous deoxythymidines in the 3â€² end of the primer; at least two non-deoxythymidine nucleotides 3â€² of the contiguous deoxythymidines; and a restriction enzyme sequence, which restriction enzyme sequence can be cleaved by a restriction enzyme when the primer is paired with its complement to provide a double-stranded restriction enzyme sequence; reverse transcribing the mRNA to produce a DNA strand complementary to the mRNA; substantially removing the polyT tail from the DNA strand; inserting the DNA strand uni-directionally into a cloning vector, and amplifying the DNA strand using a DNA polymerase; wherein a uni-directionally cloned complementary DNA library is produced.