Temperature reference plate, useful for determining temperature distribution in heating block of PCR device, comprises recesses for receiving samples having carrier plate, and cover member, where recesses are filled with reference solution

摘要

The temperature reference plate comprises: recesses for receiving samples having a carrier plate, where the recesses of the carrier plate are filled with a reference solution, reference fragments and a dye; and a cover member. The reference fragment is a double-stranded nucleic acid molecule with a known melting point, at which the double-stranded nucleic acid molecule is melted into two single stranded nucleic acid molecules. The dye comprises different binding properties compared to the double-stranded nucleic acid molecule and the single-stranded nucleic acid molecules. The temperature reference plate comprises: recesses for receiving samples having a carrier plate, where the recesses of the carrier plate are filled with a reference solution, reference fragments and a dye; and a cover member. The reference fragment is a double-stranded nucleic acid molecule with a known melting point, at which the double-stranded nucleic acid molecule is melted into two single stranded nucleic acid molecules. The dye: comprises different binding properties compared to the double-stranded nucleic acid molecule and the single-stranded nucleic acid molecules; is intercalated in the double-stranded reference fragment that is released during the melting of the double-stranded reference fragment; does not bind to the single stranded nucleic acid molecules; and is a fluorescent dye, which fluoresces in the bonded state to the double-stranded reference fragment and does not fluoresce in the unbound state. The cover member is arranged on the surface of the carrier plate in such a way that the reference solution can not escape from the respective recesses filled with the reference solution. The reference solution comprises a single-stranded interference nucleic acid molecule, whose sequence is partially complementary to the sequence of one of the strands of the reference fragment. The reference fragments exhibit different melting points. The reference fragment has a length of 200-300 or 350-460 base pairs. The melting point of the reference fragment is 85-95[deg] C. Independent claims are included for: (1) a method of determining the temperature distribution in a heating block of a PCR device; and (2) a computer program product having a computer program.