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, comprising:
a) providing:
i) a first element comprising a surface;
ii) a second element comprising bottom and top surfaces, said bottom surface comprising a plurality of indentations;
iii) a first solution comprising a plurality of first PCR primers;
iv) a second solution comprising a plurality of second PCR primers; and
v) a third solution comprising template at a known concentration;
b) contacting the surface of said first element with said first solution under conditions such that at least a portion of said first PCR primers attach to at least a portion of said surface so as to create a first modified surface comprising attached molecules;
c) contacting the bottom surface of said second element with said second solution under conditions such that at least a portion of said second PCR primers attach to at least a portion of said bottom surface so as to create a second modified surface comprising attached molecules; and
d) positioning said second modified surface, with said second PCR primers attached thereto, on top of said first modified surface, with said first PCR primers attached thereto, so as to create a plurality of first regions defined by said indentations, said first regions comprising unmasked attached molecules, and second regions comprising masked attached molecules, thereby making an array.