Affymetrix, Inc.

Quantum Dot Corporation

Vincent E Phillips

Andrew Watson

Anthony J Berno

Ghassan Ghandour

Huu Minh Tran

Thomas B Ryder

Jennifer Lai

Methods, compositions and articles of manufacture for assaying a sample for an amplification product from a target polynucleotide are provided. An amplification reaction is used to produce the amplification product from the target polynucleotide so that it can be used to indirectly assay the sample for the target polynucleotide. A sample suspected of containing the target polynucleotide is contacted with first and second primers to amplify the target polynucleotide; the first primer comprises a tag sequence, the complement of which is formed on the opposite strand during amplification and is referred to as a capture sequence. That opposite strand is referred to as a second primer extension product or an amplification product, and comprises a label. A capture probe is provided that is conjugated to a substrate and can bind to the capture sequence to form an amplification product detection complex. Methods of detecting the amplification product thus produced are also provided, as are amplification product assay arrays, along with methods of forming the same. The methods are particularly useful in multiplex settings where a plurality of target polynucleotides are to be assayed. Kits comprising reagents for performing such methods are also provided.

Methods and compositions for polynucleotide analysis using generic capture sequences

A genetic sample is analyzed by providing a gene probe array including a plurality of genetic probes having different receptors. The sample is processed to include at least one fluorescently tagged ligand. The array is hybridized by exposing the probes to the processed sample such that ligands can bind to complementary receptors. Composite data having a data dynamic range is obtained from the array using an optical scanner which has a scanner dynamic range that is smaller than the data dynamic range. The scanner optically irradiates and scans the probes and detects fluorescent emissions at a first wavelength which is selected such that the scanner produces valid first data in a low intensity portion of the data dynamic range and is in saturation in at least part of a high intensity portion of the data dynamic range. The scanner then optically irradiates and scans the probes with light and detects fluorescent emissions at a second wavelength which is selected such that the scanner produces valid second data in a high intensity portion of the data dynamic range and is in cutoff in at least part of a low intensity portion of the data dynamic range. A scale factor correlation function is calculated between the first data and the second data, and is applied to convert the second data to have a same scale factor as the first data. The first data for the first portion of the data dynamic range is combined with the converted second data for the second portion of the data dynamic range to obtain the composite data.

Method for scanning gene probe array to produce data having dynamic range that exceeds that of scanner

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Sunnyvale, CA, United States of America

Vincent E Phillips