Compositions and methods for cDNA synthesis
First Claim
1. 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/μ
l;
(c) an effective amount of oligo(dT), wherein said oligo(dT) is present in a concentration less than about 2 μ
M.
2 Assignments
0 Petitions
Accused Products
Abstract
Methods for making eDNA molecules, for amplification of RNA by PCR and for preparation of cDNA libraries are provided. Kits for making eDNA 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 eDNA and RT PCR without dilution or addition of further components. These compositions are useful, alone or in the form of kits, for eDNA 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.
55 Citations
15 Claims
-
1. 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/μ
l;(c) an effective amount of oligo(dT), wherein said oligo(dT) is present in a concentration less than about 2 μ
M. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15)
-
Specification