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US 2019/0169683 A1Published application
Detection of an Amplification Product using pH-sensitive Dye
AssigneeNew England Biolabs, Inc.
InventorsYinhua Zhang, Nathan Tanner, Thomas C. Evans, Jr.
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Abstract
Methods are provided for a rapid, low cost approach to monitoring an amplification reaction. This includes monitoring the progress of isothermal or PCR amplification reactions to completion using pH-sensitive dyes that are either colored or fluorescent. Compositions are described that include a mixture of a DNA polymerase, deoxyribonucleotide triphosphate and Tris buffer in the range of 1.5 mM Tris to 5 mM Tris or equivalent.
Claims17
An amplification method comprising:
- (a) setting up a buffered amplification reaction that comprises 1.5 mM to 5 mM Tris or an equivalent thereof and, additionally, a pH sensitive dye;
- (b) incubating the amplification reaction under amplification conditions;
- and,
- (c) after or during the incubation of (b), detecting a change in color of the amplification reaction, wherein a change in color of the reaction indicates that an amplification product has been produced.
1 more element
An aqueous amplification reaction mix comprising:
- (a) a polymerase,
- (b) 1.5 mM to 5 mM Tris or an equivalent thereof,
- (c) a single pH sensitive dye,
- (d) dNTPs,
- (e) one or more primers, and
- (f) one or more templates;
- wherein the reaction mix changes color only during the amplification reaction.
4 more elements
Dependencies are inferred from the claim text.