Methods of using dyes in association with nucleic acid staining or detection

摘要

Methods of using dyes and associated technology are provided. A dye, such as a monomeric dye or a dimeric dye, may be used in a nucleic acid gel staining application and/or a nucleic acid detection application. A dimeric dye, such as a dimeric dye capable of forming a hairpin-like structure, may be used to stain and/or detect nucleic acids via a release-on-demand mechanism. A dimeric dye having low background fluorescence in the absence of nucleic acids and high fluorescence in the presence of nucleic acids, upon binding therewith, may be used to stain and/or detect nucleic acids. Buffers comprising a weak acid and a salt of the weak acid, such as a lithium salt, are also provided to allow for effective prestaining of nucleic acids.

主权项

1. A method of detecting the presence or absence of a nucleic acid in a sample, comprising:
combining a sample comprising a nucleic acid with at least one fluorescent nucleic acid binding dye having the formula: wherein BRIDGE is a substantially neutral covalent linker comprising from about 1 to about 150 non-hydrogen atoms; Q1 is a phenanthridinium dye, an acridinium dye or an asymmetric cyanine dye; Q2 is a phenanthridinium dye, an acridinium dye or an asymmetric cyanine dye; loading said sample into a gel, wherein said gel comprising a gel buffer; immersing said gel in a running buffer; wherein said gel buffer or running buffer comprises at least one weak acid and a lithium salt of said at least one weak acid; electrophoresing said sample; and detecting fluorescence associated with the sample or a lack thereof.