US 12,247,251 B2
Enzymatic circuits for molecular sensors
Barry L. Merriman, La Jolla, CA (US); Venkatesh Alagarswamy Govindaraj, San Diego, CA (US); Paul Mola, San Diego, CA (US); and Tim Geiser, San Diego, CA (US)
Assigned to Semicon Bio, San Diego, CA (US)
Filed by Roswell Biotechnologies, Inc., San Diego, CA (US)
Filed on Dec. 30, 2020, as Appl. No. 17/138,314.
Application 16/878,484 is a division of application No. 16/684,338, filed on Nov. 14, 2019, granted, now 11,268,123.
Application 17/138,314 is a continuation of application No. 16/878,484, filed on May 19, 2020, granted, now 10,913,966.
Application 16/684,338 is a continuation of application No. 16/015,028, filed on Jun. 21, 2018, granted, now 10,508,296, issued on Dec. 17, 2019.
Application 16/015,028 is a continuation of application No. PCT/US2018/029382, filed on Apr. 25, 2018.
Claims priority of provisional application 62/489,881, filed on Apr. 25, 2017.
Prior Publication US 2022/0002777 A1, Jan. 6, 2022
This patent is subject to a terminal disclaimer.
Int. Cl. C12Q 1/6869 (2018.01); C07H 21/02 (2006.01); C12Q 1/00 (2006.01); C12Q 1/26 (2006.01); C12Q 1/48 (2006.01); C12Q 1/6825 (2018.01); G01N 33/543 (2006.01)
CPC C12Q 1/6869 (2013.01) [C07H 21/02 (2013.01); C12Q 1/001 (2013.01); C12Q 1/002 (2013.01); C12Q 1/005 (2013.01); C12Q 1/26 (2013.01); C12Q 1/48 (2013.01); C12Q 1/485 (2013.01); C12Q 1/6825 (2013.01); G01N 33/5438 (2013.01); G01N 2333/9126 (2013.01)] 15 Claims
 
1. A method to obtain a sequence information of a target substrate, the method comprising:
forming at least one of a voltage or a current circuit, the circuit further comprising:
a first electrode;
a second electrode spaced apart from the first electrode;
a first arm molecule having a first end and a second end, the second end of the first arm molecule electrically coupled to the first electrode;
a second arm molecule having a first end and a second end, the second end of the second arm molecule electrically coupled to the second electrode; and
a polymerase enzyme,
wherein the first end of each of the first and second arm molecules are conjugated to two distinct sites on the polymerase enzyme so that a portion of the polymerase enzyme between the two distinct sites is included in the circuit;
exposing the circuit to a solution having an ionic strength from dissolved ions and the target substrate;
measuring electrical signals through the circuit as the polymerase enzyme engages the target substrate; and
processing the electrical signals to identify the sequence information of the target substrate when engaged by the polymerase enzyme;
wherein the target substrate comprises DNA or RNA.