Circular chromosome conformation capture (4c)

一种染色体、核苷酸序列的技术，应用在环状染色体构象捕获(4C)领域
CN101688237AInactive Publication Date: 2010-03-31ERASMUS UNIV MEDICAL CENT ROTTERDAM ERASMUS MC
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Patent Information

Authority / Receiving Office
CN · China
Patent Type
Applications(China)
Current Assignee / Owner
ERASMUS UNIV MEDICAL CENT ROTTERDAM ERASMUS MC
Publication Date
2010-03-31
Estimated Expiration
Not applicable · inactive patent

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Abstract

The present invention relates in one aspect to a method for analysing the frequency of interaction of a target nucleotide sequence with one or more nucleotide sequences of interest (eg. one or more genomic loci) comprising the steps of: (a) providing a sample of cross-linked DNA; (b) digesting the cross-linked DNA with a primary restriction enzyme; (c) ligating the cross-linked nucleotide sequences; (d) reversing the cross linking; (e) optionally digesting the nucleotide sequences with a secondary restriction enzyme; (f) optionally ligating one or more DNA sequences of known nucleotide composition to the available secondary restriction enzyme digestion site(s) that flank the one or more nucleotide sequences of interest; (g) amplifying the one or more nucleotide sequences of interest usingat least two oligonucleotide primers, wherein each primer hybridises to the DNA sequences that flank the nucleotide sequences of interest; (h) hybridising the amplified sequence(s) to an array; and (i) determining the frequency of interaction between the DNA sequences.
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Description

field of invention

[0001] The present invention involves analyzing the frequency with which two or more nucleotide sequences interact in nuclear space. Alterations in interactions are used as a means of detecting genomic rearrangements for diagnosis and prediction. Background of the invention

[0002] The study of mammalian nuclear structure aims to understand how 2-meter-long DNA folds into a 10-μm-diameter nucleus while allowing the precise expression of genes for a given cell type and how they faithfully proliferate during each cell cycle. Much of the progress in this field has come from microscopy studies that have revealed that genomes are non-randomly arranged in nuclear space. For example, tightly packed heterochromatin is segregated from more expanded euchromatin, and chromosomes occupy different regions in nuclear space 2 . There is a complex relationship between nuclear localization and transcriptional activity. While transcription occurs throughout the interior...

Claims

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