Method for synthesizing cDNA

Abstract

A method for synthesizing cDNA characterized by performing a reverse transcription reaction in the presence of an enzyme having a reverse transcriptional activity and another enzyme different from the former one which as a 3′-5′ exonuclease activity.

1 . A method for synthesizing a cDNA, characterized in that the method comprises conducting a reverse transcription reaction in the presence of an enzyme having a reverse transcription activity and an α-type DNA polymerase having a 3′-5′ exonuclease activity.  
     
     
         2 . The method according to  claim 1 , wherein the enzyme having a reverse transcription activity is a heat-resistant reverse transcriptase.  
     
     
         3 . The method according to  claim 2 , wherein the enzyme having a reverse transcription activity is a DNA polymerase from a thermophilic bacterium of the genus  Bacillus.    
     
     
         4 . The method according to  claim 3 , wherein the enzyme having a reverse transcription activity is a DNA polymerase from  Bacillus caldotenax  or a DNA polymerase from  Bacillus stearothermophilus.    
     
     
         5 . The method according to  claim 1 , wherein the α-type DNA polymerase having a 3′-5′ exonuclease activity is an α-type DNA polymerase from a thermophilic archaebacterium.  
     
     
         6 . A method for amplifying a cDNA, comprising conducting a gene amplification reaction using a cDNA synthesized according to the method defined by  claim 1  as a template.  
     
     
         7 . The method according to  claim 6 , wherein the gene amplification reaction is a PCR.  
     
     
         8 . The method according to  claim 7 , wherein a DNA polymerase for the PCR is the same enzyme as the enzyme having a 31-5′ exonuclease activity.  
     
     
         9 . The method according to  claim 8 , wherein the DNA polymerase for the PCR is an α-type DNA polymerase from an archaebacterium.