Patent ID: 7510829

Claim:
A method of amplifying a nucleic acid template, comprising the steps of: (a) contacting a linear amplification molecule to a nucleic acid template under conditions that support specific hybridization between one end of the linear amplification molecule and the template, the linear amplification molecule having a homology region and a cleavable site, wherein said homology region and said cleavable site are between a first primer region and a second primer region, wherein said cleavable site is located 3â€² of said homology region, and wherein said homology region is between 15 and 100 nucleotides in length and has the same sequence as a sequence in the nucleic acid template, (b) adding sequentially to said linear amplification molecule at the end hybridized to said template, nucleotides complementary to corresponding nucleotides present in said template, whereby an extended linear amplification molecule containing a newly-added sequence capable of specific hybridization to the homology region in the linear amplification molecule is formed, (c) incubating said extended linear amplification molecule under conditions that support its specific intramolecular hybridization, whereby a double-stranded intramolecular hybridization region is formed between the homology region and the newly-added sequence capable of specific hybridization to the homology region, (d) cleaving said extended linear amplification molecule at the cleavable site, whereby a first fragment and a second fragment of said extended linear amplification molecule are formed, wherein said first fragment comprises said first primer region and said second fragment comprises said second primer region, and whereby said first fragment contacts said second fragment along said intramolecular hybridization region; (e) adding sequentially to said first fragment at the end newly created by said cleavage step one or more nucleotides complementary to corresponding nucleotides present in said second fragment, whereby an extended first fragment of the linear amplification molecule is formed; and (f) amplifying the extended first fragment of the linear amplification molecule by primers specific for the first and second primer regions to amplify the nucleic acid template.