Solution reagent kit MOLgen

for real-time PCRclinicalsingle nucleotide polymorphism

Reagent kit is designed for determination of single-nucleotide polymorphism of MCM6 gene - 13910C/T by real-time PCR with detection of melting curves. The kits contain reagents for real-time PCR only. Introduction “MOLgen Genetics MCM6 Kit” is intended for the determination of single nucleotide polymorphism -13910 C/T in MCM6 gene (regulatory site of LCT gene, encoding the lactase) in clinical specimens (whole blood, buccal epithelium) using the method of real-time polymerase chain reaction (PCR) with melting curve analysis. The extraction of DNA from clinical specimens can be performed usingthe “MOLgen Universal Extraction Kit”manufactured by Adaltis S.r.l. The results of PCR analysis are taken into account in complex diagnostics of disease. The kitis intended for use with block-type PCR cyclers: AMPLIlab (Adaltis), CFX96 (Bio-Rad, USA). The kit contains reagents required for 48 tests, including control samples. Principle Of Method The principle of the method is based on the amplification of the selected human DNA region with subsequent formation of hybrid duplexes of PCR product with the specific DNA probe and the detection of duplexes melting curves. The amplification step consists of repeated cycles: temperature denaturation of the DNA template, primers annealing to specific sites of the DNA template, and primer extension with Taq polymerase. After the amplification step, the temperature is lowered and the specific fluorescent-labelled DNA probe hybridizes with the quencher-labelled amplicon resulting in the decrease of the fluorescent signal. During the melting step, the DNA probe is released leading to the fluorescence increase.