US 10,889,852 B2
Genome methylation analysis
Peter Molloy, Chatswood (AU); Susan Margaret Mitchell, Gladesville (AU); and Konsta Duesing, Campbell (AU)
Assigned to Commonwealth Scientific and Industrial Research Organisation, Act (AU)
Appl. No. 15/519,893
Filed by Commonwealth Scientific and Industrial Research Organisation, Campbell (AU)
PCT Filed Oct. 20, 2015, PCT No. PCT/AU2015/050646
§ 371(c)(1), (2) Date Apr. 18, 2017,
PCT Pub. No. WO2016/061624, PCT Pub. Date Apr. 28, 2016.
Claims priority of application No. 2014904189 (AU), filed on Oct. 20, 2014.
Prior Publication US 2017/0247748 A1, Aug. 31, 2017
Int. Cl. C12Q 1/6809 (2018.01); C12Q 1/6869 (2018.01); C12Q 1/6806 (2018.01); C12Q 1/6883 (2018.01); G16B 30/00 (2019.01)
CPC C12Q 1/6809 (2013.01) [C12Q 1/6806 (2013.01); C12Q 1/6869 (2013.01); C12Q 1/6883 (2013.01); G16B 30/00 (2019.02); C12Q 2600/118 (2013.01); C12Q 2600/154 (2013.01)] 16 Claims
OG exemplary drawing
 
1. A method for identifying a site or region of a gene or genome for differential methylation analysis, the method comprising:
(a) obtaining one or more test samples of genomic DNA and one or more reference samples of genomic DNA;
(b) digesting said samples of genomic DNA obtained in (a) with a methylation-dependent restriction enzyme selected from GlaI, PcsI, KroI and combinations thereof;
(c) selecting restriction fragments generated in (b) of between about 40 bp and 500 bp;
(d) constructing libraries of said selected restriction fragments;
(e) sequencing said libraries;
(f) aligning said sequences obtained in (e) and derived from said samples of genomic DNA with a reference genome;
(g) comparing sequences consisting of said sequences aligned in (f); and
(h) identifying a methylated or unmethylated site or region of a gene or genome within the test sample of genomic DNA from said comparing of sequences.