Method of reverse transcription

Abstract

The present invention relates to reverse transcription of RNA, and in particular to reverse transcription by thermostable DNA polymerases. Thermoactinomyces vulgaris and Bacillus stearothermophilus possess reverse transcriptase activity in the presence of magnesium or manganese ions. Methods, compositions, and kits for reverse transcription and RT-PCR are also provided.

1 - 23 . (canceled) 
     
     
         24 . A kit for reverse transcription comprising:
 a) a polymerase selected from  Thermoactinomyces vulgaris  and  Bacillus stearothermophilus  polymerase,   b) purified RNA, and   c) a buffer comprising magnesium ions wherein said buffer is substantially free of manganese ions.   
     
     
         25 . The kit of  claim 24 , further comprising instructions for reverse transcription. 
     
     
         26 . The kit of  claim 24 , wherein said buffer further comprises a surfactant. 
     
     
         27 . The kit of  claim 24 , wherein said buffer has a pH of about 6 to 10. 
     
     
         28 - 29 . (canceled) 
     
     
         30 . The kit of  claim 24 , further comprising at least one additional thermostable DNA polymerase. 
     
     
         31 . A kit for reverse transcription comprising:
 a) a polymerase selected from  Thermoactinomyces vulgaris  and  Bacillus stearothermophilus  polymerase, and   b) a buffer comprising magnesium ions wherein said buffer is substantially free of manganese ions.   
     
     
         32 - 33 . (canceled) 
     
     
         34 . The kit of  claim 31 , further comprising at least one additional thermostable DNA polymerase. 
     
     
         35 . The kit of  claim 31 , further comprising instructions for reverse transcription.