US 11,692,224 B2
Synthetic nucleic acid spike-ins
Fred C. Christians, Los Altos Hills, CA (US); Igor D. Vilfan, East Palo Alto, CA (US); Michael Kertesz, Menlo Park, CA (US); Timothy A. Blauwkamp, Palo Alto, CA (US); Shivkumar Venkatasubrahmanyam, San Jose, CA (US); Michael Rosen, Palo Alto, CA (US); and Rene Sit, Sunnyvale, CA (US)
Assigned to Karius, Inc., Redwood City, CA (US)
Filed by Karius, Inc., Redwood City, CA (US)
Filed on Jun. 23, 2021, as Appl. No. 17/355,882.
Application 17/355,882 is a continuation of application No. 15/953,822, filed on Apr. 16, 2018, granted, now 11,078,532.
Application 15/953,822 is a continuation of application No. 15/469,474, filed on Mar. 24, 2017, granted, now 9,976,181, issued on May 22, 2018.
Claims priority of provisional application 62/451,363, filed on Jan. 27, 2017.
Claims priority of provisional application 62/397,873, filed on Sep. 21, 2016.
Claims priority of provisional application 62/313,668, filed on Mar. 25, 2016.
Prior Publication US 2021/0324467 A1, Oct. 21, 2021
This patent is subject to a terminal disclaimer.
Int. Cl. C12Q 1/68 (2018.01); C12Q 1/6874 (2018.01); G16B 30/00 (2019.01); C12Q 1/6869 (2018.01); G16B 30/10 (2019.01); C12N 15/10 (2006.01); C12Q 1/689 (2018.01)
CPC C12Q 1/6874 (2013.01) [C12N 15/1065 (2013.01); C12Q 1/689 (2013.01); C12Q 1/6869 (2013.01); G16B 30/00 (2019.02); G16B 30/10 (2019.02); C12Q 2600/166 (2013.01)] 49 Claims
OG exemplary drawing
 
1. A method of determining abundance of nucleic acids in an initial sample comprising target nucleic acids, the method comprising:
(a) adding a known quantity of spike-in nucleic acids to the initial sample comprising target nucleic acids, wherein the spike-in nucleic acids comprise at least 1,000 spike-in nucleic acids with sequences that are unique to each other;
(b) performing a sequencing assay on a portion of the target nucleic acids and a portion of the at least 1000 spike-in nucleic acids, thereby obtaining target nucleic acid sequence reads and spike-in nucleic acid sequence reads; and
(c) using the spike-in nucleic acid sequence reads to calculate abundance of the target nucleic acids.