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Chemically-sensitive field effect transistor based pixel array with protection diodes

a protection diode and field effect transistor technology, applied in the field of chemically sensitive field effect transistors based pixel arrays with protection diodes, can solve the problems of affecting the detection efficiency of a single sample, the inability to predict the operation of the transmission gate, and the inability to completely exclude the operation, so as to increase the number of template nucleic acids used per sensor, and optionally per reaction chamber, and increase the magnitude of charge change. , the effect of increasing the signal

Inactive Publication Date: 2013-01-17
LIFE TECH CORP
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

This patent talks about methods and compositions to improve the detection of hydrogen ions released during a nucleic acid synthesis reaction. By reducing the buffering capacity of the reaction solution or solid supports, a greater proportion of the hydrogen ions are detected by the chemFET. Additionally, the patent talks about a way to introduce a stable reference potential into a flow cell, which is not affected by changes in fluid composition or pH. These improvements make the process more reliable and accurate.

Problems solved by technology

In particular, because the body and source of the p-channel ISFET are electrically coupled together, implementing the p-channel MOSFET S11p in the same n-well as the p-channel ISFET 50 would lead to unpredictable operation of the transmission gate, or preclude operation entirely.
Analysis of entire genomes of viruses, bacteria, fungi, animals and plants is now possible, but such analysis generally is limited due to the cost and time required to sequence such large genomes.
Moreover, present conventional sequencing methods are limited in terms of their accuracy, the length of individual templates that can be sequenced, and the rate of sequence determination.

Method used

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  • Chemically-sensitive field effect transistor based pixel array with protection diodes
  • Chemically-sensitive field effect transistor based pixel array with protection diodes
  • Chemically-sensitive field effect transistor based pixel array with protection diodes

Examples

Experimental program
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Effect test

example 1

Bead Preparation

[0720]Binding of Single-Stranded Oligonucleotides to Streptavidin-Coated Magnetic Beads. Single-stranded DNA oligonucleotide templates with a 5′ Dual Biotin tag (HPLC purified), and a 20-base universal primer were ordered from IDT (Integrated DNA Technologies, Coralville, Ind.). Templates were 60 bases in length, and were designed to include 20 bases at the 3′ end that were complementary to the 20-base primer (Table 4, italics). The lyophilized and biotinylated templates and primer were re-suspended in TE buffer (10 mM Tris-HCl, 1 mM EDTA, pH 8) as 40 μM stock solutions and as a 400 μM stock solution, respectively, and stored at −20° C. until use.

[0721]For each template, 60 μl of magnetic 5.91 μm (Bangs Laboratories, Inc. Fishers, Ind.) streptavidin-coated beads, stored as an aqueous, buffered suspension (8.57×104 beads / μL), at 4° C., were prepared by washing with 120 μl bead wash buffer three times and then incubating with templates 1, 2, 3 and 4 (T1, T2, T3, T4: Ta...

example 2

On-Chip Polymerase Extension Detected by pH Shift on an ISFET Array

[0742]Streptavidin-coated 2.8 micron beads carrying biotinylated synthetic template to which sequencing primers and T4 DNA polymerase are bound were subjected to three sequential flows of each of the four nucleotides. The template sequence downstream of the sequencing primer was a G(C)10(A) 10 (SEQ ID NO:5). Each nucleotide cycle consisted of flows of dATP, dCTP, dGTP and dTTP, each interspersed with a wash flow of buffer only. Flows from the first cycle are shown in blue, flows from the second cycle in red, and the third cycle in yellow. As shown in FIG. 72A, signal generated for both of the two dATP flows were very similar. FIG. 72B shows that the first (blue) trace of dCTP is higher than the dCTP flows from subsequent cycles, corresponding to the flow in which the polymerase should incorporate a single nucleotide per template molecule. FIG. 72C shows that the first (blue) trace of dGTP is approximately 6 counts hi...

example 3

Sequencing in a Closed System and Data Manipulation

[0743]Sequence has been obtained from a 23-mer synthetic oligonucleotide and a 25-mer PCR product oligonucleotide. The oligonucleotides were attached to beads which were then loaded into individual wells on a chip having 1.55 million sensors in a 1348×1152 array having a 5.1 micron pitch (38400 sensors per mm2) About 1 million copies of the synthetic oligonucleotide were loaded per bead, and about 300000 to 600000 copies of the PCR product were loaded per bead. A cycle of 4 nucleotides through and over the array was 2 minutes long. Nucleotides were used at a concentration of 50 micromolar each. Polymerase was the only enzyme used in the process. Data were collected at 32 frames per second.

[0744]FIG. 73A depicts the raw data measured directly from an ISFET for the synthetic oligonucleotide (SE ID NO:6). One millivolt is equivalent to 68 counts. The data are sampled at each sensor on the chip (1550200 sensors on a 314 chip) many times...

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Abstract

Methods and apparatus relating to 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.

Description

CROSS-REFERENCE TO RELATED APPLICATIONS[0001]This application is a continuation of U.S. patent application Ser. No. 13 / 149,279 filed 31 May 2011, which is a continuation of U.S. patent application Ser. No. 12 / 475,311, filed 29 May 2009; the entire contents of each of which are incorporated herein by reference.REFERENCE TO BIOLOGICAL SEQUENCE DISCLOSURE[0002]This application contains nucleotide sequence and / or amino acid sequence disclosure in computer readable form and a written sequence listing, the entire contents of both of which are expressly incorporated by reference in their entirety as though fully set forth herein.FIELD OF THE DISCLOSURE[0003]The present disclosure is directed generally to inventive methods and apparatus relating to detection and measurement of one or more analytes including analytes associated with or resulting from a nucleic acid synthesis reaction.BACKGROUND[0004]Electronic devices and components have found numerous applications in chemistry and biology (...

Claims

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Application Information

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Patent Type & Authority Applications(United States)
IPC IPC(8): H01L21/336
CPCG01N27/27G01N27/4145Y10T29/49002G01N27/414C12Q1/6874C12Q1/6869G01N27/4148G01N33/54373
Inventor MILGREW, MARKBUSTILLO, JAMESREARICK, TODD
Owner LIFE TECH CORP
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