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US 2003/0143593 A1Published application
Quantative method for measuring gene expression
OwnerFabienne Hermitte, Vincent Fertinventors
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Abstract
The invention concerns a method for quantitative measuring of gene expression by obtaining marked probes of pre-selected homogeneous size, after reverse transcription and amplification. The invention also concerns the method for preparing marked probes, and implementing kits using microarrays and macroarrays.
Claims19
A process for producing probes that are representative of a population of nucleic acids the elements of which are to be analysed quantitatively, characterized in that it comprises:
- a) a step for calibrating the experimental conditions for transcription or reverse transcription to obtain nucleic acid fragments with a homogeneous size;
- b) a step for producing a population of probe sequences from transcription or reverse transcription of the population of nucleic acids the elements of which are to be quantitatively analysed, under incubation time conditions that have been preestablished during the previous step, so that the probes are of homogeneous size and are representative of the 3′ portion of each element of said population;
- c) a step for amplification of the sequences obtained at b).
A kit for the quantitative study of the variability of a transcriptome, characterized in that it comprises at least:
- free dNTP, one of which is labeled;
- a reverse transcriptase;
- at least two quantitative validation controls constituted by two sequences of nucleic acids with a predetermined size and each being different, which do not form part of the transcriptome or are not hybridizable with the elements of the transcriptome;
- a support on which are fixed in an ordered way target sequences that can be hybridized with the 3′ copies of the mRNA of transcriptome, and at least two targets that are hybridizable with the external standards.
1 more element
Dependencies are inferred from the claim text.