Method for improved reverse transcription at high temperatures

In a method for reverse transcribing RNA into DNA comprising contacting the RNA with a reverse transcriptase, wherein the improvement comprises performing said contacting in the presence of a chaperone protein, whereby the reverse transcriptase has enhanced thermostability and substantially reduced RNase H activity.

A method for enhancing thermal stability and substantially reducing RNase H activity of a reverse transcriptase comprising employing said reverse transcriptase, in a reverse transcription reaction of RNA to DNA, in the presence of a chaperone protein.

A method for reverse transcribing RNA into DNA comprising contacting the RNA with a reverse transcriptase, and a chaperone protein at a temperature above 42° C., whereby the reverse transcriptase has substantially reduced RNase H activity.

In a method for reverse transcribing RNA into DNA comprising contacting the RNA with a reverse transcriptase, wherein the improvement comprises performing said contacting in the presence of a chaperone protein, whereby the reverse transcriptase has substantially reduced RNase H activity.

A method for substantially reducing RNase H activity of a reverse transcriptase comprising employing said reverse transcriptase, in a reverse transcription reaction of RNA to DNA, in the presence of a chaperone protein.

A method for preparing DNA from RNA comprising contacting the RNA with a reverse transcriptase, wherein the contacting is in the presence of a chaperone protein, whereby the reverse transcriptase has substantially reduced RNase H activity and enhanced thermostability.

A method for preparing DNA from RNA comprising contacting the RNA with a reverse transcriptase, wherein the contacting is in the presence of a chaperone protein, whereby the reverse transcriptase has substantially reduced RNase H activity.

A method for reverse transcribing RNA to DNA comprising contacting the RNA with a reverse transcriptase, and a chaperone protein, whereby the reverse transcriptase has enhanced thermostability and substantially reduced RNase H activity.

A method for reverse transcribing RNA into DNA comprising contacting the RNA with a reverse transcriptase, and a chaperone protein, whereby the reverse transcriptase has substantially reduced RNase H activity.

A composition comprising CpkB and a reverse transcriptase.

A kit for the preparation of cDNA comprising a first container containing CpkB and a second container containing a reverse transcriptase;

• or a container containing CpkB and a reverse transcriptase mixture.

A composition for reverse transcription of a target ribonucleic acid (RNA) comprising a single lyophilizate comprising:

• a) an effective amount of a reverse transcriptase;
• b) CpkB;
• c) deoxyribonucleotide triphosphates and ribonucleotide triphosphates, wherein when said lyophilizate is reconstituted by addition of an aqueous solvent, and the resulting solution will amplify a single-stranded RNA molecule having a target nucleotide sequence region when contacted with one or more suitable oligonucleotide primers under appropriate nucleic acid amplification conditions.