Methods of RNA amplification
AssigneeAsuragen, Inc.
InventorsGary J. Latham, Richard Andrew Blidner, Liangjing Chen
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Abstract
The invention relates to methods of RNA amplification, including methods for the reverse transcription of cDNA from RNA using a thermostable reverse transcriptase. In a particular aspect, the methods are capable of linear amplification of an RNA template through multiple cycles of cDNA synthesis.
Claims23
A kit for improving cDNA synthesis by linear amplification of an RNA template, said kit comprising:
- a reaction volume containing buffer, dNTPs, betaine or a betaine analog in an amount of about 0.75 to about 3.5 M, and a first primer complementary to an RNA template;
- and
- a thermostable reverse transcriptase capable of synthesizing cDNA by linear amplification of the RNA template;
- wherein at least one cycle of cDNA synthesis includes an extension temperature and a denaturation temperature;
- and
- wherein the denaturation temperature is about 75 degrees C. to 90 degrees C.
The kit of any preceding claim, wherein the thermostable reverse transcriptase is selected from a retroviral reverse transcriptase, a family A DNA polymerase with reverse transcriptase activity, a family B DNA polymerase with reverse transcriptase activity, or a modified, mutated, or engineered variation thereof.
Dependencies are inferred from the claim text.