US 11,674,133 B2
Methods and compositions for extracting nucleic acids using ferric oxide particles
Eric P. Dixon, Cary, NC (US); John Joseph Harrington, Clayton, NC (US); Yutao Chen, Durham, NC (US); Nikhil Rao, San Francisco, CA (US); and Ling Wang, Cary, NC (US)
Assigned to BECTON, DICKINSON AND COMPANY, Franklin Lakes, NJ (US)
Appl. No. 16/644,744
Filed by Becton, Dickinson and Company, Franklin Lakes, NJ (US)
PCT Filed Sep. 10, 2018, PCT No. PCT/US2018/050157
§ 371(c)(1), (2) Date Mar. 5, 2020,
PCT Pub. No. WO2019/055331, PCT Pub. Date Mar. 21, 2019.
Claims priority of provisional application 62/558,074, filed on Sep. 13, 2017.
Prior Publication US 2020/0216830 A1, Jul. 9, 2020
Int. Cl. C12N 15/10 (2006.01); C12Q 1/6876 (2018.01)
CPC C12N 15/1013 (2013.01) [C12Q 1/6876 (2013.01); C12Q 2521/537 (2013.01); C12Q 2600/178 (2013.01)] 10 Claims
 
1. A method of generating a sample of extracted microRNAs from an initial biological sample, the method comprising:
(a) contacting the initial biological sample comprising microRNAs with proteinase K at a temperature in a range of from 47° C. to 53° C. for a period of time in a range of from 10 to 40 minutes, to degrade proteins present in the sample, thereby producing a proteinase K treated sample;
(b) contacting the proteinase K treated sample with ferric oxide particles under acidic conditions to induce binding between the ferric oxide particles and the microRNAs;
(c) magnetically separating the microRNA bound ferric oxide particles from the proteinase K treated sample;
(d) contacting the microRNA bound ferric oxide particles with an alkaline elution buffer to release the microRNAs from the ferric oxide particles into the alkaline elution buffer, thereby generating an alkaline microRNAs sample; and
(e) magnetically separating the ferric oxide particles from the alkaline microRNA sample,
to generate a sample of extracted microRNAs.