US 2010/0304995 A1 · Arrays and Methods for Reverse Genetic Functional Analysis

Abstract

Provided are methods, kits and arrays for carrying out relative measurement of an analyte of interest in a biological sample. As specifically exemplified, there is an array of stabilized, desiccated cDNA preparations, each at a defined location within the array, where those cDNAs were prepared from cells treated with a particular condition believed to modulate at least one gene of interest. Detection can be via Real Time Polymerase Chain Reaction using an appropriate reaction mixture and primers specific for a coding sequence of interest, and a greater relative amount of a RT PCR product from a control preparation reflects greater gene expression in response to the test condition whereas a lower amount of RT PCR product reflects an inhibitory effect on expression of the coding sequence of interest as a result of the application of the test condition.

Claims26

Dependent claims

A kit comprising:

a) an array comprising, at defined locations, two or more stabilized biological samples comprising an analyte of interest, wherein the samples are desiccated;
and
b) a mixture of reagents capable of detecting the analyte of interest in a container separate from the array.

A method for detecting modulation of gene expression in a cell of interest in response to a test condition, said method comprising the steps of:

a) selecting a panel of treatments, optionally from 4 to 384;
b) incubating the cell of interest in parallel under test conditions effecting the panel of treatments for a time sufficient to allow modulation of gene expression in response to at least one treatment within the panel of treatments, and optionally further comprising incubating a cell of interest in parallel with at least one negative control condition;
c) isolating a biological sample comprising an analyte of interest from the cells after step b;
d) dispensing the biological samples of step c at indexed positions of an array vessel;
e) immobilizing and/or stabilizing the biological samples in the array vessel;
f) optionally storing and optionally distributing the array of samples;
g) applying to the array a mixture of reagents capable of detecting the analyte of interest within the biological sample in an analysis reaction;
and
h) measuring output of the analysis reaction of step g,
whereby modulation of expression of a gene of interest is determined when the output of the analysis reaction for the analyte of interest is different in the biological sample prepared from cells incubated under the test conditions than in a biological sample from control cells not treated with a test condition which modulates expression of the gene of interest.

7 more elements

Dependencies are inferred from the claim text.

Arrays and Methods for Reverse Genetic Functional Analysis

Life of this patent

Arrays and Methods for Reverse Genetic Functional Analysis

Life of this patent

Abstract

Claims26