PTH Factor II PCR Assay
The PTH 20210G>A RealFast Assay detects the 20210G>A mutation in the human Factor II gene (F2), prothrombin, by real-time PCR. The mutation raises the prothrombin level in blood and thereby the risk of venous thromboembolism. Identifying carriers supports the assessment of thrombophilia risk.
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This kit in detail
- F2 20210G>A (prothrombin) genotyping
- TaqMan 5′ nuclease assay on a 110 bp fragment of the F2 gene
- Allele-specific probes: HEX for wild type, FAM for the variant
- Wild-type (GG) and homozygous mutant (AA) controls included
- HotStart Taq DNA polymerase in a ready-to-use 2x genotyping mix
- 32 or 100 reactions per kit
The PTH 20210G>A RealFast Assay from ViennaLab Diagnostics determines the genotype of the 20210G>A mutation in the Factor II gene (F2), also known as prothrombin.
Clinical background
Thrombophilia is an increased tendency to form blood clots. The 20210G>A mutation in F2 leads to an elevated prothrombin level in blood, which raises the risk of thrombotic events and of venous thromboembolism. Identifying carriers provides a basis for risk assessment and for preventive measures.
Method
The assay uses the TaqMan principle, a fluorogenic 5′ nuclease technique. The RealFast 2x Genotyping Mix contains HotStart Taq DNA polymerase and dNTPs in an optimised buffer; the assay mix contains F2-specific primers that amplify a 110 bp fragment of the F2 gene, together with two allele-specific, dual-labelled hydrolysis probes.
Read-out
During PCR the exonuclease activity of the Taq polymerase cleaves the fluorescent reporter from the hybridised probe, and the signal is measured in real time. In a wild-type sample the HEX-labelled probe binds, giving a strong HEX signal and little FAM signal; in a homozygous mutant sample the FAM-labelled probe binds, giving the reverse pattern; a heterozygous sample gives an intermediate signal in both channels.
Controls and format
A wild-type control (GG) and a homozygous mutant control (AA) are supplied. The kit is available in 32 and 100 reaction formats.
- RealFast™ 2x Genotyping Mix: Contains HotStart Taq DNA polymerase and dNTPs in an optimized buffer system for PCR amplification.
- PTH 20210G>A Assay Mix: Includes F2 gene-specific primers and two allele-specific, dual-labeled hydrolysis probes for genotyping.
- PTH 20210G>A WT-Control: A control sample representing the wild-type genotype (GG).
- PTH 20210G>A MUT-Control: A control sample representing the homozygous mutant genotype (AA).
Datasheets and regulatory documentation
Everything your quality team needs for evaluation and procurement.
Instructions for Use
Frequently paired with this kit
Other assays and instruments lab teams add alongside the PTH Factor II PCR Assay.
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