Agarose droplet microfluidics for highly parallel and efficient single molecule emulsion PCR.
An agarose droplet method was developed for highly parallel and efficient single molecule emulsion PCR. The method capitalizes on the unique thermoresponsive sol-gel switching property of agarose for highly efficient DNA amplification and amplicon trapping. Uniform agarose solution droplets generated via a microfluidic chip serve as robust and inert nanolitre PCR reactors for single copy DNA molecule amplification. After PCR, agarose droplets are gelated to form agarose beads, trapping all amplicons in each reactor to maintain the monoclonality of each droplet. This method does not require cocapsulation of primer labeled microbeads, allows high throughput generation of uniform droplets and enables high PCR efficiency, making it a promising platform for many single copy genetic studies.
Leng
Xuefei
X
State Key Laboratory of Physical Chemistry of Solid Surfaces, The Key Laboratory for Chemical Biology of Fujian Province, Key Laboratory of Analytical Sciences, and Department of Chemical Biology, College of Chemistry and Chemical Engineering, Xiamen University, Xiamen, 361005, P.R.China.
Zhang
Wenhua
W
Wang
Chunming
C
Cui
Liang
L
Yang
Chaoyong James
CJ
D004655:Emulsions; D016133:Polymerase Chain Reaction / Q000379:methods*; D012685:Sepharose
D004655:Emulsions; D012685:Sepharose