Primer middle sequence interference pcr technology

A method for inhibiting primer dimer formation and non-specific amplification of a target sequence therefrom in a polymerase chain reaction (PCR), the method comprising amplifying the target sequence using:

• (a) a first primer and a second primer which, when aligned in the 5′ to 3′ direction, share an identical Intermediate Domain (ID) sequence of 5-9 consecutive bases, wherein both primers specifically bind and amplify the target sequence while primer dimer formation and non-specific amplification therefrom are inhibited;
• (b) a first primer, a second primer, and an oligonucleotide which is complementary to an Intermediate Domain (ID) sequence of 5-11 consecutive bases within the first primer or the second primer, wherein the oligonucleotide (As Oligo) does not affect the primers specifically binding and amplifying the target sequence while inhibiting primer dimer formation and non-specific amplification therefrom;
• and/or (c) a first primer and a second primer, wherein the first primer or second primer comprises an Intermediate Domain (ID) sequence of 5-7 consecutive bases and an anti-sense sequence complementary to the ID sequence, wherein the anti-sense sequence is capable of intra-molecular binding to the ID sequence such that the primer is folded, and wherein both the first and second primers specifically bind and amplify the target sequence while primer dimer formation and non-specific amplification therefrom are inhibited.