Solution reagent kit ME138020

for real-time PCRclinicalgenetic

MOLgen Genetics Haemostatic Disease (F2/F5) reagent kit is intended for differential determination of single-nucleotide polymorphisms of the coagulation system and folate cycle by real-time PCR with detection of melting curves. The kits contain reagents for real-time PCR only. Introduction “Molgen Genetics Haemostatic Disease (F2/F5) Kit” is intended for differential determination of polymorphisms 20210 G/A of F2 gene (coagulation factor II, prothrombin) and 1691 G/A (Leiden mutation) of F5 gene (coagulation factor V, proaccelerin) by real-time polymerase chain reaction (PCR) followed by melting of hybridisation products with a fluorescent tag and detection of melting curves. The kit is intended for use with block-type PCR cycler CFX96 (Bio-Rad, USA) such as Adaltis AmpliLab. The assay kit can be applied in clinical practice for a comprehensive study of the genetic factors which increase the risk of disorders in the blood coagulation system. The extraction of human DNA from clinical samples (buccal epithelium, whole blood) is performed using the kit “MOLgen Universal Extraction Kit”. The kit is designed for the analysis of 48 samples, including control samples. Principle Of Method The principle of the method is based on the amplification of the selected human DNA region followed by detection of melting curves of hybrid complexes in PCR products and specific probes. The process of amplification consists in repeated cycles: temperature denaturation of DNA matrix, annealing primers with complementary sequences of DNA matrix, and synthesis of a complementary chain from these primers with Taq polymerase.