Reaction mixture for positioning a reaction vessel relative to a detection unit

Abstract

The present invention concerns methods for measuring a nucleic acid amplification in real-time comprising (i) providing a real-time PCR instrument containing a reaction vessel holder for holding multiple reaction vessels and means for positioning the multiple reaction vessels in a reaction vessel holder relative to the detection unit, (ii) filling multiple reaction vessels with a reaction mixture containing thermostable polymerase, deoxynucleotides and buffer, at least two amplification primers, at least one fluorescently-labelled hybridization probe and a fluorescent dye component which is present in a free form during and after the entire amplification reaction, (iii) determining the position of maximum fluorescence emission of the fluorescent dye component in multiple reaction vessels as a function of their position relative to the detection unit, and (iv) performing the amplification reaction and measuring the fluorescence emission of the at least one hybridization probe during and optionally after the amplification reaction for each reaction vessel at the position of maximum fluorescence emission.

Dependencies are inferred from the claim text.