Patent ID: 8354233

Claim:
A method for reducing the background signal generated during a nucleic acid sequencing reaction, the method comprising the steps of: (a) amplifying a nucleic acid by an amplification reaction with a forward and a reverse primer, wherein the forward or the reverse primer is phosphorylated on the 5′ end; (b) performing a chain termination reaction in the presence of said forward and reverse amplification primers with a sequencing primer to generate primer extension products incorporating at least one additional nucleotide, wherein said sequencing primer is not phosphorylated on the 5′ end; (c) after the chain termination reaction, enzymatically degrading chain termination extension products having said phosphorylated primer incorporated thereto using a nuclease having specificity for a 5′-phosphorylated substrate; and (d) determining the polynucleotide sequence of the amplified nucleic acid.