METHODS AND APPARATUS FOR MEASURING ANALYTES

• US 20130217004A1
• Filed: 09/12/2012
• Published: 08/22/2013
• Est. Priority Date: 10/22/2008
• Status: Abandoned Application

• Current Assignee: Life Technologies Corporation (Thermo Fisher Scientific Incorporated)
• Original Assignee: Life Technologies Corporation (Thermo Fisher Scientific Incorporated)
• Inventors: ROTHBERG, Jonathan M., Hinz, Wolfgang, Davidson, John F., van Oijen, Antoine M., Huber, Martin, Leamon, John H.
• Application Number:
  US13/612,742
• Publication Number:
  US 20130217004A1
• Time in Patent Office: Days
• US Class Current: 435/6.1
• CPC Class Codes: C12Q 1/6869 Methods for sequencing
• CPC Class Codes: C12Q 1/6874 involving nucleic acid arrays, e.g. sequencing by hybridisation
• CPC Class Codes: C12Q 2565/607 being a sensor, e.g. electrode
• CPC Class Codes: G01N 27/4145 specially adapted for biomolecules, e.g. gate electrode with immobilised receptors
• CPC Class Codes: G01N 27/4148 Integrated circuits therefor, e.g. fabricated by CMOS processing CMOS processing per se H01L21/82
• CPC Class Codes: H01L 21/82 to produce devices, e.g. integrated circuits, each consisting of a plurality of components

Abstract

Methods and apparatus relating to FET arrays including large FET arrays for monitoring chemical and/or biological reactions such as nucleic acid sequencing-by-synthesis reactions. Some methods provided herein relate to improving signal (and also signal to noise ratio) from released hydrogen ions during nucleic acid sequencing reactions.

Claims

1. A method for sequencing a nucleic acid comprising:

contacting and incorporating known nucleotides into a plurality of identical nucleic acids in a reaction chamber in contact with or capacitively coupled to an ISFET, wherein the nucleic acids are covalently bound to a single bead in the reaction chamber, and

detecting hydrogen ions released upon nucleotide incorporation in the presence of no or limited buffering activity.

2. The method of claim 1, wherein the ISFET is in an ISFET array that comprises 256 ISFET.

3. The method of claim 1, wherein the reaction chamber comprises a solution having no buffer or low buffer concentration.

4. The method of claim 1, wherein the reaction chamber comprises a solution having a buffering inhibitor.

5. The method of claim 1, wherein the single bead is at least 90% saturated with nucleic acids.

6. The method of claim 1, wherein the nucleic acids are sequencing primers.

7. The method of claim 6, wherein the nucleic acids are hybridized to template nucleic acids.

8. The method of claim 6, wherein the nucleic acids are hybridized to concatemers of identical template nucleic acids.

9. The method of claim 1, wherein the nucleic acids are self-priming template nucleic acids.

10-154. -154. (canceled)