A method of quantitating an amount of a target DNA template in a sample comprising: a) forming a plurality of mixture partitions, wherein the mixture partitions comprise: i) the target DNA template, wherein the target DNA template is less than 90 nucleotides in length; ii) a thermostable DNA dependent DNA polymerase; and iii) a forward and a reverse amplification primer, wherein the amplification primers comprise a 3' hybridization region that hybridizes to the target DNA template and primes template directed extension of the primer in the presence of the DNA dependent DNA polymerase; the forward and/or the reverse amplification primer further comprises a 5' tail region that is not complementary to the target DNA template; and the forward and reverse amplification primers have a combined length greater than the length of the target DNA template; and b) incubating the mixture partitions under thermal cycling conditions suitable for amplification of the target DNA template by a polymerase chain reaction, wherein the thermal cycling conditions comprise a first set of temperature cycles and a second set of temperature cycles, wherein the first set of temperature cycles comprises an annealing temperature of less than 50 °C, and the second set of temperature cycles comprises an annealing temperature of at least 50 °C and which is at least 5 °C higher than an annealing temperature of the first set of temperature cycles; and c) detecting the presence or absence of amplified target DNA template in the mixture partitions and determining the fraction of partitions where the target DNA template is present; thereby quantifying the amount of target DNA template in the sample. a) forming a plurality of mixture partitions, wherein the mixture partitions comprise: i) the target DNA template, wherein the target DNA template is less than 90 nucleotides in length; ii) a thermostable DNA dependent DNA polymerase; and iii) a forward and a reverse amplification primer, wherein the amplification primers comprise a 3' hybridization region that hybridizes to the target DNA template and primes template directed extension of the primer in the presence of the DNA dependent DNA polymerase; the forward and/or the reverse amplification primer further comprises a 5' tail region that is not complementary to the target DNA template; and the forward and reverse amplification primers have a combined length greater than the length of the target DNA template; and i) the target DNA template, wherein the target DNA template is less than 90 nucleotides in length; ii) a thermostable DNA dependent DNA polymerase; and iii) a forward and a reverse amplification primer, wherein the amplification primers comprise a 3' hybridization region that hybridizes to the target DNA template and primes template directed extension of the primer in the presence of the DNA dependent DNA polymerase; the forward and/or the reverse amplification primer further comprises a 5' tail region that is not complementary to the target DNA template; and the forward and reverse amplification primers have a combined length greater than the length of the target DNA template; and the amplification primers comprise a 3' hybridization region that hybridizes to the target DNA template and primes template directed extension of the primer in the presence of the DNA dependent DNA polymerase; the forward and/or the reverse amplification primer further comprises a 5' tail region that is not complementary to the target DNA template; and the forward and reverse amplification primers have a combined length greater than the length of the target DNA template; and b) incubating the mixture partitions under thermal cycling conditions suitable for amplification of the target DNA template by a polymerase chain reaction, wherein the thermal cycling conditions comprise a first set of temperature cycles and a second set of temperature cycles, wherein the first set of temperature cycles comprises an annealing temperature of less than 50 °C, and the second set of temperature cycles comprises an annealing temperature of at least 50 °C and which is at least 5 °C higher than an annealing temperature of the first set of temperature cycles; and c) detecting the presence or absence of amplified target DNA template in the mixture partitions and determining the fraction of partitions where the target DNA template is present; thereby quantifying the amount of target DNA template in the sample.