公开(公告)号：US11155792B2

公开(公告)日：2021-10-26

申请号：US16789278

申请日：2020-02-12

Applicant: ARIZONA BOARD OF REGENTS ON BEHALF OF ARIZONA STATE UNIVERSITY

Inventor： John Chaput ,  Andrew Larsen ,  Matthew Dunn

IPC: C12N9/12 ,  C12Q1/68 ,  G01N21/64 ,  B01L3/00

Abstract: The present invention is directed to a polymerase activity assay that produces a strong optical signal when a primer-template complex is extended to full-length product. The assay uses Cy3 as the molecular beacon and Iowa Black® RQ as the quencher. The signal-to-noise-ratio (STNR) of this donor-quencher pairing is ˜200-fold over background, which is considerably better than other donor-quencher pairs (STNRs ˜10-20-fold). The STNR allows for solution-based monitoring of polymerase activity. Because the sensor functions via Watson-Crick base pairing, the polymerase activity assay may also be used to evolve polymerases to accept xeno nucleic acids as substrates.

公开(公告)号：US10584319B2

公开(公告)日：2020-03-10

申请号：US15766716

申请日：2016-10-10

Applicant: ARIZONA BOARD OF REGENTS ON BEHALF OF ARIZONA STATE UNIVERSITY

Inventor： John Chaput ,  Andrew Larsen ,  Matthew Dunn

IPC: C12N9/12 ,  G01N21/64 ,  C12Q1/68 ,  B01L3/00

Abstract: The present invention is directed to a polymerase activity assay that produces a strong optical signal when a primer-template complex is extended to full-length product. The assay uses Cy3 as the molecular beacon and Iowa Black® RQ as the quencher. The signal-to-noise-ratio (STNR) of this donor-quencher pairing is ˜200-fold over background, which is considerably better than other donor-quencher pairs (STNRs ˜10-20-fold). The STNR allows for solution-based monitoring of polymerase activity. Because the sensor functions via Watson-Crick base pairing, the polymerase activity assay may also be used to evolve polymerases to accept xeno nucleic acids as substrates.

公开(公告)号：US20180320150A1

公开(公告)日：2018-11-08

申请号：US15766716

申请日：2016-10-10

Applicant: ARIZONA BOARD OF REGENTS ON BEHALF OF ARIZONA STATE UNIVERSITY

Inventor： John Chaput ,  Andrew Larsen ,  Matthew Dunn

IPC: C12N9/12 ,  C12Q1/68 ,  G01N21/64

CPC classification number: C12N9/1252 ,  B01L3/5027 ,  C12N9/1241 ,  C12Q1/68 ,  C12Y207/07007 ,  G01N21/64

Abstract: The present invention is directed to a polymerase activity assay that produces a strong optical signal when a primer-template complex is extended to full-length product. The assay uses Cy3 as the molecular beacon and Iowa Black® RQ as the quencher. The signal-to-noise-ratio (STNR) of this donor-quencher pairing is ˜200-fold over background, which is considerably better than other donor-quencher pairs (STNRs ˜10-20-fold). The STNR allows for solution-based monitoring of polymerase activity. Because the sensor functions via Watson-Crick base pairing, the polymerase activity assay may also be used to evolve polymerases to accept xeno nucleic acids as substrates.