Patent ID: 11946043
Assignee: LUDWIG INSTITUTE FOR CANCER RESEARCH LTD
Field: Biotechnology (Chemistry)
Classification: CPC C | IPC C

Claim 0:
1. A method for creating a library of 5-methylcytosine (5mC) containing DNA from a low-input DNA sample comprising the steps in order of:
(a) obtaining a low-input DNA sample;
(b) adding a blocking group to the endogenous 5-hydroxymethylcytosine (5hmC) in the DNA sample;
(c) converting the 5mC in the DNA sample to 5hmC with Naeglerai gruberi TET1 (NgTET1);
(d) adding a linking group to the 5hmC from step (c);
(e) after step (d), ligating DNA adapters to the DNA sample;
(f) after step (e), adding an affinity tag to the linking group from step (d);
(g) enriching for the affinity tagged DNA from step (f) by affinity purification utilizing a support; and
(h) amplifying the enriched DNA from step (g), wherein amplifying the enriched DNA comprises PCR directly on the enriched DNA linked to the support.