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US 2014/0273100 A1Published application
cDNA SYNTHESIS METHOD
AssigneeSeiko Epson Corporation
InventorsYuji Saito, Fumio Takagi
Life of this patent
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Abstract
A cDNA synthesis method includes: mixing a lysis solution containing a chaotropic substance and a nucleic acid-binding solid-phase carrier in a sample containing a ribonucleic acid (RNA), thereby adsorbing the RNA on the carrier; reverse-transcribing the RNA adsorbed on the carrier while keeping the RNA adsorbed on the carrier in a reverse transcription reaction mixture, thereby synthesizing cDNA; and eluting the synthesized cDNA with an eluent.
Claims9
A cDNA synthesis method, comprising:
- mixing a lysis solution containing a chaotropic substance and a nucleic acid-binding solid-phase carrier in a sample containing a ribonucleic acid (RNA), thereby adsorbing the RNA on the carrier;
- reverse-transcribing the RNA adsorbed on the carrier while keeping the RNA adsorbed on the carrier in a reverse transcription reaction mixture, thereby synthesizing cDNA;
- and
- eluting the synthesized cDNA with an eluent.
1 more element
A cDNA synthesis kit, comprising:
- a neutral lysis solution containing 4 to 7 M of a guanidine salt, 0 to 5% of a nonionic surfactant, and 0 to 0.2 M of a reducing agent;
- a nucleic acid-binding solid-phase carrier;
- a first washing solution containing 4 to 7 M of a guanidine salt and 0 to 5% of a nonionic surfactant;
- a second washing solution composed of water or a low salt concentration aqueous solution;
- a reverse transcription reaction mixture containing a reverse transcriptase, dNTP, and a primer for reverse transcription;
- and
- a DNA amplification mixture containing a DNA polymerase, dNTP, and primers for DNA amplification.
4 more elements
Dependencies are inferred from the claim text.