DETERMINATION OF VARIANTS PRODUCED UPON REPLICATION OR TRANSCRIPTION OF NUCLEIC ACID SEQUENCES

摘要

A method of determining whether or not a nucleic acid having an expected sequence or one or more variants of the expected sequence are present in a sample containing nucleic acids after replication, transcription or editing (or other transformation) of a substrate nucleic acid. The method involves deciding an expected sequence likely to be formed in the sample upon the replication, transcription or editing of the substrate nucleic acid, and possible variants of the expected sequence, providing primer pairs for a polymerase chain reaction, reverse transcriptase polymerase chain reaction or ligase chain reaction, carrying out the polymerase chain reaction or reverse transcriptase polymerase chain reaction in one or more steps to form amplicons, and analyzing the amplicons to determining whether or not a nucleic acid having the expected sequence and/or variants are present in the sample. The primers of the primer pairs are designed to anneal to regions of the nucleic acid of the expected sequence and the variants, the regions being selected to reveal unambiguously the presence or absence in the sample of the nucleic acid of the expected sequence or the variants thereof according to the presence or absence of specific amplicons amplified by the primers.

主权项

1. A method of determining whether or not a nucleic acid having an expected sequence or one or more variant nucleic acids having a variant of the expected sequence is or are present in a sample containing nucleic acids after replication, transcription or editing of a substrate nucleic acid, which method comprises:
deciding an expected sequence likely to be formed in said sample upon said replication, transcription or editing of said substrate nucleic acid, and possible variants of said expected sequence; providing at least one primer pair for amplifying a nucleic acid sequence in a nucleic acid amplification step, carrying out said amplification step to form one or more amplicons, and analyzing said one or more amplicons to determining whether or not a nucleic acid having said expected sequence or one or more variants are present in said sample, wherein primers of said at least one primer pair are designed to anneal to regions of said nucleic acid of said expected sequence and said one or more variant nucleic acid sequences, said regions being selected to reveal unambiguously the presence or absence in said sample of said nucleic acid of said expected sequence or said one or more variants thereof according to the presence or absence of amplicons amplified by said primers.