US 11,913,062 B2
System and method for isolation and qualification of nucleic acids
Victoria Constandse, Cape Town (ZA); Daniel Klass, Livermore, CA (US); Alexander Lovejoy, Newark, CA (US); Melissa Loyzer, Hamilton (CA); Bronwen Miller, Cape Town (ZA); Bernd Hinzmann, Berlin (DE); and Sophie Beckert, Neuzelle (DE)
Assigned to Roche Sequencing Solutions, Inc., Pleasanton, CA (US); and Signature Diagnostics GmbH, Potsdam (DE)
Filed by Roche Sequencing Solutions, Inc., Pleasanton, CA (US)
Filed on Nov. 18, 2019, as Appl. No. 16/687,274.
Application 16/687,274 is a continuation of application No. PCT/US2018/033279, filed on May 17, 2018.
Claims priority of provisional application 62/507,501, filed on May 17, 2017.
Prior Publication US 2020/0087708 A1, Mar. 19, 2020
Int. Cl. C12Q 1/68 (2018.01); C12Q 1/6806 (2018.01); C40B 20/08 (2006.01); C40B 30/10 (2006.01); C40B 60/06 (2006.01); G16B 40/10 (2019.01); G16B 30/00 (2019.01)
CPC C12Q 1/6806 (2013.01) [C40B 20/08 (2013.01); C40B 30/10 (2013.01); C40B 60/06 (2013.01); G16B 40/10 (2019.02); G16B 30/00 (2019.02)] 2 Claims
OG exemplary drawing
 
1. A method comprising:
(a) isolating DNA from a source, thereby providing a composition comprising the isolated DNA, wherein the isolated DNA has at least a first target region having a first length in nucleotides and a second target region having a second length in nucleotides, wherein the second length is greater than the first length, and the first target region and the second target region are a long interspersed nuclear element (LINE);
(b) quantifying a total mass of the isolated DNA in the composition;
(c) quantifying a first concentration of the first target region and a second concentration of the second target region in the composition;
(d) calculating a Q-ratio for the isolated DNA by dividing the second concentration by the first concentration;
(e) determining a value for a quality-mass constant (kQm) by a method comprising: (i) sequencing a plurality of regions of a plurality of isolated DNA samples, wherein each of the isolated DNA samples has a known input mass and a known Q-ratio; (ii) comparing a fraction (FD) of the plurality of the regions with a selected depth of coverage as determined by sequencing in step (a) with a product of the input mass and the Q-ratio for each of the isolated DNA samples; (iii) identifying a minimum value for the product of the input mass and the Q-ratio at which FD is greater than a target FD; and (iv) setting the quality-mass constant (kQm) equal to the minimum value for the product of the input mass and the Q-ratio determined in step (iii);
(f) estimating a required input mass for the isolated DNA by dividing the quality-mass constant (kQm) by the Q-ratio; and
(g) preparing the isolated DNA for sequencing if the total mass of the isolated DNA in the composition is equal or greater than the required input mass.