Patent ID: 6773901
Filing Date: 2004-08-10
Classification: C12Q

Abstract:
A method for determining a base sequence of a primer comprising:carrying out a PCR by using four types of primers which respectively have a structure comprising a first sequence of a given base length complementary to one of single strands of a target DNA and a second sequence of a given base length provided adjacent to the side of 5â€² terminus of said first sequence and being non-complementary to the one of single strands of the target DNA; analyzing results of amplified products obtained by the PCR; requiring efficiencies of adenylation using results of the step of analyzing; determining one out of the four types of primers as a sequence which is most likely to undergo adenylation, wherein each of the four types of primers have one base at 5â€² terminus of the second sequence, the one base being different among the four types of primers; carrying out a second PCR by using another four types of primers which respectively have the one base at the 5â€² terminus of the second sequence of the sequence determined in the step of determining one out of the four types of primers, and have a second base located at a second site from the 5â€² terminus thereof and different among the another four types of primers; analyzing second results of amplified products by the second PCR; requiring efficiencies of adenylation using results of the step of analyzing the second results; and determining one out of the another four types of primers to be most likely to undergo adenylation.