Sensing apparatus and method

Abstract

Provided is a sensing apparatus comprising a chip for integrated amplification and sequencing of a template polynucleotide in a sample. The apparatus comprises a chip with at least one ISFET in a well or chamber, amplification means for amplifying the template polynucleotide on a surface of said chip and comprising at least one heating means suitable for conducting amplification of the template polynucleotide at temperatures elevated with respect to room temperature, and sequencing means for sequencing the amplified template polynucleotide in said well or chamber. Methods of use are also provided.

Description

BACKGROUND[0001]Over the last two decades, there has been a rapid development in nucleic acid analysis, specifically in the field of nucleic acid amplification and DNA sequencing technology, with an increasing range of instrumentation now available. The conventional methods of detecting and analysing a nucleic acid sequence primarily rely on fluorescent nucleic acid intercalating dyes, fluorescent-labelled oligonucleotide probes, fluorescent- or radioactive-labelled nucleotides.[0002]Subsequently, a new method of analysing nucleic acid synthesis and sequencing has been developed using a semiconductor-based detection system such as an Ion Sensitive Field Effect Transistor (ISFET), see for instance our PCT publication WO 03 / 073088. An Ion Sensitive Field Effect Transistor -based platform, unlike conventional fluorescent-based nucleic acid analysis systems, does not require expensive optical instruments or dangerous radioactive isotopes for detection, thus making this platform a cost e...

AI Technical Summary

Benefits of technology

The patent describes a method for amplifying DNA in a well-controlled environment. During amplification, a removable seal is provided to prevent cross-contamination between wells and to contain the amplification reactions. The method also involves using an ISFET to monitor the amplification reaction and detect the presence of suitable colonies of template. This allows the system to change or stop the amplification process in a specific well or all wells. The use of the removable seal and the monitoring of the amplification reaction improve the efficiency and accuracy of the DNA amplification process.

Problems solved by technology

However, such spatial separation of the two steps requires a skilled work force, high levels of hands on time, introduces incremental errors and may increase sample loss, thus decreasing the sensitivity of detection for sequencing and increasing costs.

It does not disclose sequence detection via sequencing-by-synthesis, only detection of the amplification activity itself.

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