Split-cycle and tape amplification

A plurality of mixture partitions, the individual mixture partitions comprising:

• i) a mutation-specific 5′-tailed primer pair, wherein the mutation-specific 5′-tailed primer pair hybridizes to and specifically amplifies target DNA template molecules from a nucleic acid sample that comprise a mutant target sequence, if present, wherein the primers of the mutation-specific 5′-tailed primer pair comprise:
• a) a 3′ hybridization region that specifically hybridizes to the mutant target sequence;
• and
• b) a mutation-specific 5′ tail region that does not hybridize to any nucleic acid fragments in the nucleic acid sample;
• ii) a wild-type-specific 5′-tailed primer pair, wherein the wild-type-specific 5′-tailed primer pair hybridizes to and specifically amplifies target DNA template molecules comprising a wild-type target sequence, if present, wherein the primers of the wild-type-specific 5′-tailed primer pair comprise:
• a) a 3′ hybridization region that specifically hybridizes to the wild-type target sequence;
• and
• b) a wild-type-specific 5′ tail region that does not hybridize to any nucleic acid fragments in the nucleic acid sample, wherein the wild-type-specific 5′ tail region is a different sequence than the mutation-specific 5′ tail region;
• and
• iii) a mutation-specific flanking primer pair, wherein the mutation-specific flanking primer pair hybridizes to and specifically amplifies amplicons comprising the 5′ tail regions of the mutation-specific 5′-tailed primer pair, if present;
• iv) a wild-type-specific flanking primer pair, wherein the wild-type-specific flanking primer pair hybridizes to and specifically amplifies amplicons comprising the 5′ tail regions of the wild-type-specific 5′-tailed primer pair, if present;
• and
• v) a thermostable polymerase, wherein
• at least some of the mixture partitions of the plurality of mixture partitions contain a target DNA template molecule that comprises a wild-type or mutant target sequence;
• and
• at least some of the mixture partitions of the plurality of mixture partitions do not contain the target DNA template molecule.