Compositions and Methods for cDNA Synthesis

摘要

The methods and compositions for preparing cDNAs, and more particularly, compositions having propylene glycol 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 are provided. The reagent mixture comprises a ready to use reagent solution, wherein the solution comprises: (a) propylene glycol in a concentration between about 25% and about 50%; 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) propylene glycol in a concentration between about 25% and about 50%; 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 Avian Myeloblastosis Virus Reverse Transcriptase, Respiratory Syncytial Virus Reverse Transcriptase, Moloney Murine Leukemia Virus Reverse Transcriptase, Human Immunodeficiency Virus Reverse Transcriptase, Equine Infectious Anemia Virus Reverse Transcriptase, Rous-Associated Virus 2 Reverse Transcriptase, Avian Sarcoma Leukosis Virus Reverse Transcriptase, RNaseH (−) Reverse Transcriptase, SuperScript II Reverse Transcriptase, and ThermoScript Reverse Transcriptase,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.