Leland Stanford Junior University

Swissfluidics Ag

Reto B Schoch

Juan G Santiago

Mostafa Ronaghi

Alexandre Persat

Robert D Chambers

A method and system are presented for fast and efficient isolation, purification and quantitation of nucleic acids from complex biological samples using isotachophoresis in microchannels. In an embodiment, a sieving medium may be used to enhance selectivity. In another embodiment, PCR-friendly chemistries are used to purify nucleic acids from complex biological samples and yield nucleic acids ready for further analysis including for PCR. In another embodiment, small RNAs from biological samples are extracted, isolated, preconcentrated and quantitated using on-chip ITP with a high efficiency sieving medium. The invention enables fast concentration and separation (takes 10s to 100s of seconds) of nucleic acids with high selectivity and using lower volumes of reagents (order of 10s of &#x3bc;L to focus less than 1 &#x3bc;g/&#x3bc;L of nucleic acid).

Isotachophoretic focusing of nucleic acids

A method and system are presented for fast and efficient isolation, purification and quantitation of nucleic acids from complex biological samples using isotachophoresis in microchannels. In an embodiment, a sieving medium may be used to enhance selectivity. In another embodiment, PCR-friendly chemistries are used to purify nucleic acids from complex biological samples and yield nucleic acids ready for further analysis including for PCR. In another embodiment, small RNAs from biological samples are extracted, isolated, preconcentrated and quantitated using on-chip ITP with a high efficiency sieving medium. The invention enables fast concentration and separation (takes 10s to 100s of seconds) of nucleic acids with high selectivity and using lower volumes of reagents (order of 10s of &#x3bc;L to focus less than 1 pg/&#x3bc;L of nucleic acid).

An isotachophoresis method for preconcentrating and isolating a plurality n of charged analytes (A, with i=1 to n) contained in a sample is disclosed, wherein each one of the analytes Ahas an effective electrophoretic mobility &#x3bc;obeying the fully ordered relationship &#x3bc;&#x3e;&#x3bc;&#x3e;etc.&#x3e;&#x3bc;, comprising the step of preparing a mixture of said sample and a number n&#x2212;1 of spacer compounds (S, with k=1 to n&#x2212;1) wherein each one of said spacer compounds (S) has an effective electrophoretic mobility &#x3bc;obeying the fully ordered relationship &#x3bc;&#x3e;&#x3bc;&#x3e;&#x3bc;. An axial electric field is applied along the longitudinal axis of a separation channel, thereby causing a preconcentration and separation of the analytes and spacers forming respective focused spacer zones and focused analyte zones that flow along the longitudinal axis. Each one of the spacer compounds (S) has an initial concentration (c) selected in such manner as to substantially correspond, at its preconcentrated concentration (c) in the respective focused spacer zone, to a volume of the main separation channel enclosed between an associated pair of adjacent extraction channels (E) and (E). Through this spatial displacement of analytes Aby spacers S, the analytes can be selectively extracted into the extraction channels E.

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Speicherschwendi, Switzerland

Reto B Schoch