Linear Amplification of Short Nucleic Acids

摘要

The present teachings provide novel methods for amplifying short nucleic acids. In some embodiments, the present teachings provide novel methods for linearly amplifying a collection of micro RNAs by using temperature cycling during a reverse transcription reaction. The cycling can comprise at least 20 cycles of an annealing temperature segment of 10° C.-30° C., and a denaturation temperature segment of 35° C.-60° C. In some embodiments, the temperature cycled reaction can comprise an osmolyte.

主权项

1. A method of linearly amplifying a multiplicity of different short nucleic acid sequences, wherein the multiplicity of short nucleic acid sequences are 15-35 nucleotides in length, said method comprising;
forming a reaction mixture comprising a multiplicity of different short nucleic acids, a multiplicity of target -specific primers, and an enzyme that catalyzes target-specific primer extension; annealing the collection of target-specific primers to the multiplicity of different short nucleic acids; extending the multiplicity of target-specific primers with the enzyme that catalyzes target-specific primer extension; cycling the reaction mixture for at least 20 cycles between an annealing temperature segment of 10° C.-30° C., and a denaturation temperature segment of 35° C.-60° C.; and, linearly amplifying the multiplicity of different short nucleic acid sequences.