Compounds for use in PCR systems and applications thereof
This disclosure relates to novel compounds for use in various compositions, kits and methods, including, for example, use in polymerase storage buffers and in nucleic acid synthesis or amplification reactions such as a polymerase chain reaction (PCR). Methods for preparing the novel compounds are also described.
1. A compound, having the structure:
wherein each n is independently any positive integer, including but not limited to 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, and 30.
2. A method for inhibiting inactivation of a polymerase in a thermal cycling process, said method comprising contacting said polymerase with the compound of claim 1 during the thermal cycling process.
3. The method of claim 2 , wherein said inhibition of polymerase inactivation is the same as the inhibition of polymerase inactivation when the polymerase is in the presence of NP-40 and/or Tween® 20 instead of said compound.
4. The method of claim 2 , wherein said inhibition of polymerase inactivation is greater than the inhibition of polymerase inactivation when the polymerase is in the presence of NP-40 or Tween® 20 instead of said compound.
5. The method of claim 2 , wherein said polymerase is thermostable.
6. The method of claim 2 , wherein said 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.
7. The method of claim 5 , wherein said 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.
8. The method of claim 7 , wherein said thermostable polymerase is Taq DNA polymerase.
9. The method of claim 7 , wherein said thermostable polymerase is Tfl DNA polymerase.
10. A method for producing a modified detergent comprising:
a) activating BrijL-23 to produce BrijL-23-Mesolate;
b) preparing BrijL-23-Iodide from BrijL-23-Mesolate;
c) preparing BrijL-23-Proline-t-Butyl ester from BrijL-23-Iodide; and,
d) hydrolyzing BrijL-23-Proline-t-Butyl ester to produce BrijL-23-Proline;
wherein said modified detergent comprises the compound of claim 1 .
11. The method of claim 10 , further comprising neutralizing the product.
12. The method of claim 11 , wherein said product is neutralized using Amberlite®.