Compositions and Methods for cDNA Synthesis

摘要

The present invention relates to methods and compositions for preparing cDNAs, and more particularly, compositions having trehalose for synthesizing a cDNA molecule or molecules from an mRNA template or population of mRNA templates under conditions sufficient to increase the detection sensitivity and cDNA yield and to simplify and improve the reliability of reverse transcription. 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%; and (b) a viral reverse transcriptase 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, in a buffer suitable for use in a reverse transcription reaction, wherein the buffer further 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%; and (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,in a buffer suitable for use in a reverse transcription reaction, wherein the buffer further comprises:
a co-factor metal ion necessary for reverse transcriptase activity; and nucleoside triphosphates.