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1. (WO1997008185) USE OF A 5' POSITIONAL PRIMER TO PRODUCE DOUBLE-STRANDED DNA FROM A DNA TEMPLATE

Pub. No.:    WO/1997/008185    International Application No.:    PCT/US1996/014046
Publication Date: Mar 6, 1997 International Filing Date: Aug 26, 1996
IPC: C12N 15/10
C12Q 1/68
Applicants: ALPHAGENE, INC.
Inventors: LEE, Ike, W.
MADIGAN, Sanford, J.
SCHEELE, George
Title: USE OF A 5' POSITIONAL PRIMER TO PRODUCE DOUBLE-STRANDED DNA FROM A DNA TEMPLATE
Abstract:
Methods for making double-stranded DNA from a linear DNA template including the steps of (i) adding a 3' tail to a strand of DNA, (ii) hybridizing a primer to the tailed DNA, which primer is complementary to the 3' tail and to a portion of the DNA, and (iii) synthesizing a second strand of DNA are disclosed, such as the method described in the figure. The methods can be used to produce ds DNA molecules having blunt ends or DNA molecules having ss DNA extensions which facilitate subsequent cloning of the DNA. Also disclosed are methods for comparing the compositions of two populations of RNA molecules, and for identifying a gene which is differentially expressed in two cell types.