Methods and devices for amplification of nucleic acid
The present invention relates to methods and devices for amplifying nucleic acid, and, in particular, amplifying so as to generate products on a surface without the use of emulsions. In a preferred embodiment, a plurality of groups of amplified product are generated on the surface, each group positioned in different (typically predetermined) locations on said surface so as to create an array.
1. A method of amplifying and sequencing nucleic acid, comprising:
a) providing i) a population of different nucleic acid template molecules, ii) a plurality of first and second single stranded oligonucleotides immobilized on a glass surface, iii) amplification reagents, iv) sequencing reagents, and v) a plurality of first and second sequencing primers;
b) hybridizing at least a portion of said population of nucleic acid template molecules to said plurality of first oligonucleotides immobilized on said glass surface;
c) amplifying said nucleic acid template molecules so as to create a plurality of forward and reverse single stranded strands;
d) sequencing said forward oligonucleotide strands with said first sequencing primers; and
e) sequencing said reverse oligonucleotide strands with said second sequencing primers.
2. The method of claim 1 , wherein said amplification reagents comprise polymerase And dNTPs.
3. The method of claim 1 , wherein said sequencing reagents comprise reagents for sequencing by synthesis.
4. The method of claim 1 , wherein said plurality of first and second single stranded oligonucleotides are immobilized to said glass surface through a linker.
5. The method of claim 4 , wherein said linker elevates said oligonucleotides away from said glass surface.
6. The method of claim 1 , wherein when the first sequencing primers are used in step
d), said second sequencing primers are not active.