Patent ID: 7981617

Claim:
A method comprising: constructing, by PCR, a transcription template having a promoter base sequence for cell-free protein synthesis using two polynucleotides (A) and (B) having base sequences complementary to different parts of the promoter base sequence, wherein (i) polynucleotide (A) is a 5′-end side primer that has a base sequence complementary to a part of a base sequence at the 5′-end of the promoter base sequence and (ii) polynucleotide (B) is a 5′-end side primer that has a base sequence different from the base sequence of polynucleotide (A) and is complementary to a part of a base sequence at the 3′-end of the promoter base sequence; and (iii) a part of a base sequence at the 3′-end of polynucleotide (A) overlaps a part of a base sequence at the 5′-end of polynucleotide (B); wherein both polynucleotides (A) and (B) are used together to construct the transcription template and transcription only occurs when polynucleotides (A) and (B) are used together.