Compositions and methods for upregulation of reverse transcription and reduction of sequence bias in rna sequencing
OwnerRaj Chakrabarti, Anisha Ghosh +3inventors
InventorsRaj Chakrabarti, Anisha Ghosh, Xiangying Guan, Gauthier Errasti, Thomas Delacroix
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Abstract
Compositions and methods for the upregulation of reverse transcription (RT) reactions and the reduction of sequence bias in RNA sequencing are presented. The compositions and methods include the application of certain organic cosolvents to increase the enzymatic activity of RT enzymes above their activity in aqueous media in addition to reducing the secondary structure of RNA, and/or the introduction of amino acid mutations that render the enzymes even more active in the presence of the organic cosolvents.
Claims35
-134. (canceled)
A composition for performing a reverse transcriptase reaction comprising:
- a thermostable reverse transcriptase or a fragment thereof;
- a reverse transcriptase (RT) buffer;
- one or more template RNAs and deoxyribonucleoside triphosphates (dNTPs);
- one or more low molecular weight polar organic solvents;
- and
- optionally a DNA-dependent DNA polymerase enzyme or a fragment thereof.
Dependencies are inferred from the claim text.