Compositions and Methods for RT-PCR

摘要

The present invention relates to methods and compositions having trehalose and DNA polymerase for facilitating the rapid and efficient amplification of nucleic acid molecules and the detection and quantitation of RNA molecules, and for increasing the detection sensitivity and reliability through generation of secure cDNA molecules prior to gene-specific primer dependent amplification. The reagent mixture comprises a ready to use reagent solution, wherein the solution comprises: (a) trehalose in a concentration between about 5% and about 35%; (b) a viral reverse transcriptase; and (c) at least one DNA polymerases, in a buffer suitable for use in a reverse transcription reaction, wherein the buffer comprises a co-factor metal ion and nucleoside triphosphates.

主权项

1. A reagent mixture comprising a ready to use reagent solution, wherein the solution comprises:
(a) trehalose in a concentration between about 5% and about 35%; (b) a viral reverse transcriptase in a concentration sufficient for use in a reverse transcription reaction without adding additional reverse transcriptase, wherein the viral reverse transcriptase is selected from the group consisting of AMV RT, RSV RT, MMLV RT, HIV RT, EIAV RT, RAV2 RT, ASLV RT, RNaseH (−) RT, SuperScript II RT, and ThermoScript RT; and (c) at least one DNA polymerases selected from the group consisting of Taq, Tth, Tfi, Tbr, Tma, Tne, pfu, Pwo, Vent, Deep Vent, Kod, and mutants thereof,in a buffer suitable for use in a reverse transcription reaction, wherein the buffer comprises:
a co-factor metal ion necessary for reverse transcriptase activity; and nucleoside triphosphates.