Non-emulsion methods and masked biomolecules
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 for making an array, comprising:
a) providing:
i) a first element comprising a surface;
ii) a second element comprising bottom and top surfaces, said second element further comprising a plurality of channels extending from said bottom to said top surfaces;
iii) a third element comprising a surface,
iv) a first solution comprising a plurality of first PCR primers further comprising anchoring molecules attached thereto, and
v) a second solution comprising templates, polymerase, dNTPs and second PCR primers,
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 substantially the entire said surface via said anchoring molecule so as to create a modified surface comprising attached first PCR primers, wherein said contacting results in the entire surface being contacted with said solution, wherein said attached first PCR primers are attached at a substantially even density; and
c) positioning said second element on the surface of said first element such that the bottom surface of said second element is on top of said modified surface, so as to create first regions defined by channels, said first regions comprising unmasked attached first PCR primers, and second regions comprising masked attached first PCR primers, whereby said positioning removes at least a portion of said solution from said surface of said first element, and
d) contacting said first regions with said second solution, and
e) positioning said third element such that said second element is between the surfaces of said first and third elements, so as to create first regions defined by sealed channels, thereby making an array, and
f) thermocycling said array under conditions such that at least a portion of said templates are amplified by said unmasked attached first PCR primer and said second PCR primer.
2. The method of claim 1 , wherein said channels of said second element are evenly spaced.