Multiple displacement amplification
First Claim
1. A method comprising:
- contacting a sample of nucleic acid with a reaction mixture comprising a set of oligonucleotide primers, one or more polymerase enzymes, and one or more water-in-oil emulsion-forming components wherein said emulsion-forming components comprise polysorbate 40 and polydimethylsiloxane such that said sample and said reaction mixture form an emulsion;
subjecting said emulsion to a multiple displacement amplification reaction comprising thermocycling the emulsion, wherein a nucleic acid sequence in said sample is amplified within at least one emulsion micro-droplet to produce at least one multiple displacement amplification product; and
converting said emulsion to aqueous and hydrophobic phases upon completion of said multiple displacement amplification reaction by adding an emulsion-breaking compound that is chloroform to said reaction.
1 Assignment
0 Petitions
Accused Products
Abstract
The present invention provides methods kits and systems for performing multiple displacement amplification reactions. In one method a sample of nucleic acid is provided. The nucleic acid is contacted with a reaction mixture which includes a set of oligonucleotide primers, a one or more polymerase enzymes and a detergent. The reaction mixture is then subjected to conditions under which the nucleic acid sequence is amplified to produce an amplified product in a multiple displacement reaction. The method may also be carried out by contacting the nucleic acid with the reaction mixture in the form of an emulsion. A kit is also provided for carrying out either the methods described above. The kit includes one or more polymerases, a plurality of primers and a detergent. The kit may also include a hydrophobic polymer and may include instructions for performing a multiple displacement amplification reaction on a nucleic acid sample.
-
Citations
16 Claims
-
1. A method comprising:
-
contacting a sample of nucleic acid with a reaction mixture comprising a set of oligonucleotide primers, one or more polymerase enzymes, and one or more water-in-oil emulsion-forming components wherein said emulsion-forming components comprise polysorbate 40 and polydimethylsiloxane such that said sample and said reaction mixture form an emulsion; subjecting said emulsion to a multiple displacement amplification reaction comprising thermocycling the emulsion, wherein a nucleic acid sequence in said sample is amplified within at least one emulsion micro-droplet to produce at least one multiple displacement amplification product; and converting said emulsion to aqueous and hydrophobic phases upon completion of said multiple displacement amplification reaction by adding an emulsion-breaking compound that is chloroform to said reaction. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16)
-
Specification