Methods of rna amplification
AssigneeMeta Platforms, 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.
Claims25
1-52. (canceled)
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 OmniAmp, an RTX polymerase, Hawk-Z05, Hawk-Z05 Fast, KOD, Maxima Reverse Transcriptase, Maxima H Minus Reverse Transcriptase, MonsterScript, Pyrophage, Pyrophage 3173, RocketScript RT, SunScript reverse transcriptase RNase H+, SunScript reverse transcriptase RNase H+, Superscript IV and derivatives, Taq, TGIRT-III enzyme, ThermoScript RT, Tth, Volcano2G DNA polymerase, and Warmstart RTx Reverse Transcriptase.
Dependencies are inferred from the claim text.