Patent ID: 6929914

Claim:
A method for producing a template for nucleotide sequence determination of an unknown sequence of a target nucleic acid molecule, comprising: (a) contacting in the presence of a nucleic acid polymerase said target nucleic acid molecule with a first primer, said first primer being complementary to a first known sequence of said target nucleic acid to synthesize a first strand; (b) removing said first primer; (c) contacting said target nucleic acid molecule with a second primer, said second primer being complementary to a second known sequence of said target molecule at the 5′ end of said second primer and being random at the 3′ end of said second primer; (d) removing said second primer; (e) trimming said first strand back to a branchpoint; (f) converting the sequence at said branchpoint of said first strand to form a first strand lariat; (g) extending said lariat to complete a lariat stem to generate a template molecule, said template molecule being suitable for sequence determination wherein said method is carried out in the absence of a restriction enzyme.