Isotachophoresis for purification of nucleic acids
The present disclosure relates to fluidic systems and devices for processing, extracting, or purifying one or more analytes. These systems and devices can be used for processing samples and extracting nucleic acids, for example by isotachophoresis. In particular, the systems and related methods can allow for extraction of nucleic acids, including non-crosslinked nucleic acids, from samples such as tissue or cells. The systems and devices can also be used for multiplex parallel sample processing.
1. A microfluidic device comprising one or more fluidic paths, wherein at least one of the fluidic paths comprises:
a branched fluidic channel comprising a loading section and an isotachophoresis (“ITP”) section wherein the ITP section comprises a leading electrolyte branch and an elution branch, meeting at a branch point;
wherein:
(a) the loading section comprises:
(1) a trailing electrolyte well fluidically connected to a sample input region of the fluidic channel; and
(2) a sample input well fluidically connected to the sample input region of the fluidic channel;
(b) a first capillary barrier is positioned between the sample input region of the fluidic channel and the ITP section; and
(c) in the ITP section, (i) the leading electrolyte branch comprises a leading electrolyte well fluidically connected to the fluidic channel; and
(ii) the elution branch comprises a sample elution well fluidically connected to the fluidic channel.
2. The microfluidic device of claim 1 , wherein the loading section further comprises a trailing capillary barrier positioned between the trailing electrolyte well and the sample input well, and a gas channel connected to the trailing capillary barrier and to a pneumatic pressure port.
3. The microfluidic device of claim 1 , wherein the ITP section further comprises a leading electrolyte buffering well fluidically connected to the leading electrolyte well through a leading electrolyte buffering channel.
4. The microfluidic device of claim 3 , wherein the leading electrolyte buffering channel comprises a leading capillary barrier and a gas channel connected to the leading capillary barrier and to a pneumatic pressure port.
5. The microfluidic device of claim 1 , wherein the elution branch further comprises an elution buffering well fluidically connected to the elution well through an elution buffering channel.
6. The microfluidic device of claim 5 , wherein the elution buffering channel comprises an elution capillary barrier, and a gas channel connected to the elution capillary barrier and to a pneumatic pressure port.
7. The microfluidic device of claim 5 , wherein the elution buffering channel comprises an elution capillary barrier, and a gas channel connected to the elution capillary barrier and to a pneumatic pressure port.
8. The microfluidic device of claim 1 , wherein:
(A) the loading section further comprises a trailing capillary barrier positioned between the trailing electrolyte well and the sample input well, and a gas channel connected to the trailing capillary barrier and to a pneumatic pressure port;
(B) the ITP section further comprises a leading electrolyte buffering well fluidically connected to the leading electrolyte well through a leading electrolyte buffering channel; and
(C) the elution branch further comprises an elution buffering well fluidically connected to the elution well through an elution buffering channel.
9. The microfluidic device of claim 8 , wherein:
the leading electrolyte buffering channel comprises a leading capillary barrier and a gas channel connected to the leading capillary barrier and to a pneumatic pressure port; and
the elution branch further comprises an elution buffering well fluidically connected to the sample elution well through an elution buffering channel.
10. The microfluidic device of claim 8 , further comprising:
an electrode positioned in the elution buffering well;
an electrode positioned in the leading electrolyte buffering well; and
an electrode positioned in the trailing electrolyte well.
11. The microfluidic device of claim 1 , comprising a plurality of the at least one of the fluidic paths.
12. The microfluidic device of claim 1 , comprising, in two interlocking parts:
(A) an insert comprising the fluidic channel, the sample input well, the trailing electrolyte well, and the leading electrolyte well; and
(B) an outer ring comprising mating features to interlock with the insert.
13. The microfluidic device of claim 12 , wherein the outer ring is sized and dimensioned to conform to a microtiter plate standard.
14. The microfluidic device of claim 1 , wherein the loading section further comprises a gas channel connected to the first capillary barrier and to a pneumatic pressure port.