µParaflo® microfluidic chip (Image Credit: LC Sciences)
A requirement common for all targeted sequencing methods, whether hybridization capture based or multiplex PCR based is a need for large numbers of custom oligonucleotide primers or probes, which can reach into the tens of thousands.
LC Sciences synthesize thousands of custom oligonucleotide sequences at once in massive parallel on a microchip and then cleave the oligos, releasing them into solution in a single micro-tube ready for use.
OligoMix® synthesis is performed on the µParaflo® microfluidic synthesis platform where synthesis occurs via the same standard DMT chemistry used for conventional oligo synthesis. No specialized nucleotide monomers are required, assuring efficient stepwise yield and high quality final product. Instead, innovative PGA chemistry uses light to generate reaction reagents and thus enables the miniaturized synthesis of custom sequences with otherwise conventional chemistry.
The functionalized µParaflo® chips are particularly suited for applications where small sample consumption, contamination-free, and performance-reproducibility are primary concerns. This technology enables the massively parallel synthesis of high quality DNA and RNA oligonucleotides as well as peptides and peptidomimetics in picoliter-scale reaction chambers.
The microfluidic chip is a closed system made of individual, parallel reaction chambers manufactured on a silicon substrate and covered with glass. The reactor contains three topographical features: pico-scale reaction chambers, fluid micro-channels, and inlet/outlet holes. The fluid micro-channels are designed with a tapered shape that was derived from a fluid mechanical model to produce a uniform flow rate across all reaction chambers. This technology enables a high density of uniform spots.
Microfluidics deliver reagent precursors to the chambers, a programmed mask is digitally projected onto the microfluidic chip prior to a beam of light. The mask determines which chambers are exposed to the light and thus generate the required reaction reagent. The specific nucleotide is incorporated into the growing oligo only at these select reactors.
These digital optics and a programmable process make parallel synthesis of a large number of different molecules on the same reaction surface simple and efficient. For more info see www.lcsciences.com
- Zhou X, Cai S, Hong A, You Q, Yu P, Sheng N, Srivannavit O, Muranjan S, Rouillard JM, Xia Y. (2004) Microfluidic PicoArray synthesis of oligodeoxynucleotides and simultaneous assembling of multiple DNA sequences. Nucleic Acids Research 32(18), 5409-5417. [article]