cDNA synthesis improvements

Abstract

The present invention generally relates to methods of making cDNA molecules and cDNA libraries. The invention also relates to cDNA molecules and cDNA libraries produced according to these methods, as well as to vectors and host cells containing such cDNA molecules and libraries. The invention also relates to kits for making the cDNA molecules and libraries of the invention.

1 . (canceled) 
     
     
         2 . A composition comprising at least one polypeptide having reverse transcriptase activity and an antibody or antibody fragment inhibitor of the polypeptide having reverse transcriptase activity. 
     
     
         3 . The composition of  claim 2 , further comprising one or more mRNA templates, or one or more poly A RNA templates and a primer. 
     
     
         4 . The composition of  claim 2 , wherein said antibody or antibody fragment is monoclonal. 
     
     
         5 . The composition of  claim 2 , wherein said polypeptide is a reverse transcriptase selected from the group consisting of Moloney Murine Leukemia Virus reverse transcriptase (M-MLV RT), Rous Sarcoma Virus reverse transcriptase (RSV RT), Avian myeloblastosis Virus reverse transcriptase (AMV RT), Rous associated Virus reverse transcriptase (RAV RT), Myeloblastosis associated virus reverse transcriptase (MAV RT) and Human Immunodeficiency Virus reverse transcriptase (HIV RT), and fragments thereof having reverse transcriptase activity. 
     
     
         6 . The composition of  claim 5 , wherein said reverse transcriptase is reduced in RNase H activity. 
     
     
         7 . The composition of  claim 6 , wherein said RNase H activity is reduced to less than about 30% of RNase H activity of a corresponding wildtype reverse transcriptase. 
     
     
         8 . The composition of  claim 2 , wherein said inhibitor inhibits, prevents, or reduces internal priming. 
     
     
         9 . The composition of  claim 3 , wherein the primer to template ratio is between 12:1 and 1:12. 
     
     
         10 . The composition of  claim 9 , wherein said primer to template ratio is between 10:1 and 1:10. 
     
     
         11 . The composition of  claim 9 , wherein said primer to template ratio is between 5:1 and 1:5. 
     
     
         12 . The composition of  claim 3 , wherein said primer has a length of between 20 and 100 bases. 
     
     
         13 . The composition of  claim 12 , wherein said length is between 20 and 75 bases. 
     
     
         14 . The composition of  claim 12 , wherein said length is between 20 and 50 bases. 
     
     
         15 . The composition of  claim 12 , wherein said length is between 25 and 35 bases. 
     
     
         16 . The composition of  claim 2 , wherein said polypeptide is a retroviral reverse transcriptase. 
     
     
         17 . The composition of  claim 2 , wherein said polypeptide is a reverse transcriptase selected from the group consisting of M-MLV RT, RSV RT and AMV RT. 
     
     
         18 . The composition of  claim 17 , wherein said reverse transcriptase is a M-MLV RT having an RNase H activity less than about 30% of the RNase H activity of the corresponding wildtype M-MLV RT. 
     
     
         19 . The composition of  claim 17 , wherein said reverse transcriptase is selected from the group consisting of SUPERSCRIPT™ (mutant M-MLV RT having reduced RNase H activity), SUPERSCRIPT™ II (mutant M-MLV RT having reduced RNase H activity), THERMOSCRIPT™ (mutant AMV RT having reduced RNase H activity) and THERMOSCRIPT™ II (mutant AMV RT having reduced RNase H activity). 
     
     
         20 . The composition of  claim 2 , wherein said one or more mRNA templates is a population of mRNA templates suitable for the production of a cDNA library. 
     
     
         21 . The composition of  claim 2 , wherein said primer is an oligo(dT) primer.