NUCLERA LTD

Disclosed is a method for the cell-free expression of peptides or proteins in a liquid filled digital microfluidic device. The droplets having the components required for cell-free protein expression can be manipulated by electrokinesis in order to enhance levels of protein expression in the droplets.

Provided herein is a method for measuring the properties of biomolecules by measuring the diffusion within a merged droplet on a microfluidic device.

A method for moving an aqueous droplet comprising providing an electrokinetic device including a first substrate having a matrix of electrodes, wherein each of the matrix electrodes is coupled to a thin film transistor, and wherein the matrix electrodes are overcoated with a functional coating comprising: a dielectric layer in contact with the matrix electrodes, a conformal layer in contact with the dielectric layer, and a hydrophobic layer in contact with the conformal layer; a second substrate comprising a top electrode; a spacer disposed between the first substrate and the second substrate and defining an electrokinetic workspace; and a voltage source operatively coupled to the matrix electrodes. The method further comprises disposing an aqueous droplet on a first matrix electrode; and providing a differential electrical potential between the first matrix electrode and a second matrix electrode with the voltage source, thereby moving the aqueous droplet.

Provided herein are methods and compositions for the on-device detection of protein synthesis using fluorescent proteins. The methods are applicable to monitoring on a microfluidic device.

A method for moving an aqueous droplet comprising providing an electrokinetic device including a first substrate having a matrix of electrodes, wherein each of the matrix electrodes is coupled to a thin film transistor, and wherein the matrix electrodes are overcoated with a functional coating comprising: a dielectric layer in contact with the matrix electrodes, a conformal layer in contact with the dielectric layer, and a hydrophobic layer in contact with the conformal layer; a second substrate comprising a top electrode; a spacer disposed between the first substrate and the second substrate and defining an electrokinetic workspace; and a voltage source operatively coupled to the matrix electrodes. The method further comprises disposing an aqueous droplet on a first matrix electrode; and providing a differential electrical potential between the first matrix electrode and a second matrix electrode with the voltage source, thereby moving the aqueous droplet.

The invention provided herein relates to methods for protein synthesis, purification and characterisation.

Provided herein are linear expression constructs and methods of cell-free protein synthesis, optimised cell-free protein synthesis (CFPS) reagents, and methods for optimising CFPS reagents to increase protein expression yields. The constructs and methods are applicable to protein expression on a microfluidic device having hydrophobic surfaces. The constructs are applicable for making membrane or other hydrophobic proteins have multiple solubility tags.

Provided herein are methods for the controlled filling of multiple reservoirs on a microfluidic device with a defined area of fluid, the method comprising: a. providing a device with a plurality of wells connected via entry holes to a substrate bearing a plurality of electrodes; b. filling the device with an aqueous immiscible filler liquid such that the plurality of wells are at least partially filled with the filler liquid; c. loading two or more of the wells with an aqueous liquid which descends in the wells; and d. withdrawing a portion of the filler liquid from the device, thereby drawing the aqueous liquids onto the plurality of electrodes through the entry holes in order to form multiple differently sized volumes of aqueous liquid.

An electrowetting system for actuating droplets of a first composition and of a second composition. The system includes: a plurality of electrodes configured to manipulate droplets of fluid in a microfluidic space, each electrode being coupled to circuitry which applies driving voltages to the electrode; and a processing unit operably connected to a look up table correlating drive sequences to chemical species and at least one composition parameter. The pulse sequence of driving voltages to move at least one of the droplets contains at least three consecutive positive pulses or is positively biased.