Compositions and methods for cDNA synthesis
AssigneeQuanta Biosciences
InventorsAyoub Rashtchian, David Schuster
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Abstract
Methods for making cDNA molecules, for amplification of RNA by PCR and for preparation of cDNA libraries are provided. Kits for making cDNA molecules also are provided. Compositions are also provided comprising mixtures of reagents, including reverse transcriptases, buffers, cofactors and other components, suitable for immediate use in conversion of RNA into cDNA and RT PCR without dilution or addition of further components. These compositions are useful, alone or in the form of kits, for cDNA synthesis or nucleic acid amplification (e.g., by the Polymerase Chain Reaction) or for any procedure utilizing reverse transcriptases in a variety of research, medical, diagnostic, forensic and agricultural applications.
Claims16
A method for reverse transcription of one or more nucleic acid molecules comprising incubating one or more nucleic acid templates in a buffer under conditions sufficient to make one or more first nucleic acid molecules complementary to all or a portion of said one or more templates, wherein said buffer comprises:
- (a) at least one reverse transcriptase;
- (b) an effective amount of a mixture of random primers, wherein said random primers are present in a concentration of at least about 5 ng/μl;
- (c) an effective amount of oligo(dT), wherein said oligo(dT) is present in a concentration less than about 2 μM.
A method for reverse transcription of one or more nucleic acid molecules comprising incubating one or more nucleic acid templates in a buffer under conditions sufficient to make one or more first nucleic acid molecules complementary to all or a portion of said one or more templates, wherein said buffer comprises:
- (a) at least one reverse transcriptase;
- (b) one or more primers suitable for priming reverse transcription of said one or more templates;
- and
- (c) an effective amount of Li ion.
16-21. (canceled)
Dependencies are inferred from the claim text.