Development of Novel Detergents for Use in PCR Systems

Patent metadata

Field	Value
Publication number	US-2017073746-A1
Application number	US-201615273824-A
Country	US
Kind code	A1
Filing date	Sep 23, 2016
Priority date	Jun 8, 2011
Publication date	Mar 16, 2017
Grant date	—

Abstract

This disclosure relates to novel detergents for use in various procedures including, for example, nucleic acid amplification reactions such as polymerase chain reaction (PCR). Methods for preparing the modified detergents are also described.

First claim

1 .- 19 . (canceled)

20 . A method for detecting a target nucleic acid in a sample, said method comprising the steps of:
a) forming a reaction mixture comprising at least one polymerase, at least one primer, dNTPs, at least one compound selected from

wherein each n is independently 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30, and a detectable label;
b) amplifying said target nucleic acid; and
c) detecting a signal generated from said detectable label indicative of the presence and/or amount of said target nucleic acid in said sample.

21 .- 26 . (canceled)

27 . The method of claim 20 , wherein said polymerase is thermostable.

28 . A nucleic acid amplification reaction mixture comprising:
a) at least one polymerase; b) dNTPs; and c) at least one compound selected from

wherein each n is independently 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30.

29 . The reaction mixture according to claim 28 , wherein said polymerase is thermostable.

30 .- 32 . (canceled)

33 . A method for amplifying a target nucleic acid comprising the steps of:
a) combining the target nucleic acid with at least one polymerase and at least one compound selected from

wherein each n is independently 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30,
to form a reaction mixture; and
b) amplifying the target nucleic acid.

34 . The method of claim 33 wherein the reaction mixture further comprises at least one nucleic acid primer and dNTPs.

35 . The method of claim 33 , further comprising a step wherein the amplified target nucleic acid is detected.

36 . The method of claim 35 , wherein the amplified target nucleic acid is detected using a detectable label.

37 . The method of claim 36 , wherein the detectable label is part of a primer or a probe.

38 . The method of claim 35 , further comprising a step wherein the amplified target nucleic acid is quantitated.

39 . The method of claim 33 , wherein the polymerase is selected from the group consisting of T7 DNA polymerase, eukaryotic mitochondrial DNA Polymerase γ, prokaryotic DNA polymerase I, prokaryotic DNA polymerase II, prokaryotic DNA polymerase III, prokaryotic DNA polymerase IV, prokaryotic DNA polymerase V, eukaryotic polymerase α, eukaryotic polymerase β, eukaryotic polymerase γ, eukaryotic polymerase δ, eukaryotic polymerase ε, eukaryotic polymerase η, eukaryotic polymerase ζ, eukaryotic polymerase ι, eukaryotic polymerase κ, E. coli DNA polymerase I, E. coli DNA polymerase III alpha subunit, E. coli DNA polymerase III epsilon subunits, E. coli polymerase IV, E. coli polymerase V, T. aquaticus DNA polymerase I, B. stearothermophilus DNA polymerase I, a Euryarchaeota polymerase, terminal deoxynucleotidyl transferase (TdT), S. cerevisiae polymerase 4, a translesion synthesis polymerase, reverse transcriptase, a thermostable polymerase, and telomerase.

40 . The method of claim 39 wherein the thermostable polymerase is selected from the group consisting of Taq DNA polymerase, Tfi DNA polymerase, Tfl DNA polymerase, Pfu DNA polymerase, and Vent™ DNA polymerase, a polymerase having reduced 3′-to-5′ exonuclease activity, SuperScript™ DNA polymerase, a genetically engineered DNA polymerase, a polymerase having the active site mutation F667Y, a polymerase having the equivalent of active site F667Y, Tth polymerase, AmpliTaq®FS, ThermoSequenase™, Therminator I, Therminator II, Therminator III, Therminator Gamma, a derivative thereof, and a fragment thereof.

41 . The method of claim 40 , wherein the thermostable polymerase is Taq DNA polymerase.

42 . The method of claim 40 , wherein the thermostable polymerase is Tfl DNA polymerase.

43 .- 57 . (canceled)

Assignees

• Life Technologies Corp

Inventors

• Angrish Parul
• Yang Zhiwei
• Wang Jonathan

Classifications

• C12Q1/6876

  Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes · CPC title

• C12Q2600/158

  Expression markers · CPC title

• C12Q1/6848 Primary

  characterised by the means for preventing contamination or increasing the specificity or sensitivity of an amplification reaction · CPC title

• C07C229/12 Primary

  to carbon atoms of acyclic carbon skeletons · CPC title

• C07D211/60

  Carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals · CPC title