System and methods of using microfluidic agitation to enhance multiplexed solid-phase isothermal nucleic acid amplification

A method of producing a micropost field, wherein the method comprises the steps of:

• a. providing a micropost template, wherein the micropost template is configured for forming a field of microposts arranged on a backing substrate with micropost-free regions for printing a capture probe array;
• b. filling the micropost template with a magnetoelastomeric material, wherein filling the micropost template comprises overfilling the template thereby forming the backing substrate connecting individual microposts in the micropost field;
• c. curing the magnetoelastomeric material;
• d. releasing the backing substrate and attached microposts from the micropost template to provide a micropost field on a backing substrate;
• and
• e. performing a quality control process to assess:
• i. the features of the microposts;
• and
• ii. the response of the microposts to a magnetic field.

A method of producing a capture probe array on a substrate surface of a micropost field, wherein the method comprises the steps of:

• a. providing a micropost field on a backing substrate, wherein the micropost field and backing substrate are formed of a magnetoelastomeric material;
• b. modifying the surface of the backing substrate for binding a plurality of oligonucleotide primers;
• c. printing an array of oligonucleotide primers on the surface of the backing substrate, thereby providing an array of capture spots comprising a plurality of immobilized oligonucleotide primers, wherein the plurality of oligonucleotide primers comprises a plurality of single target specific primers per capture spot, and one or more process controls;
• d. performing a quality control process to assess:
• i. the features of the primer array;
• and
• ii. target binding.