人类线粒体DNA损伤 qPCR试剂盒

该试剂盒针对的是最容易受到损伤的长mtDNA区域（D-Loop基因）。原则上，更多的mtDNA损伤或病变可以减缓或阻断DNA聚合酶的进展，但完整的mtDNA则不会。基因特异性Taqman™探针qPCR是高度敏感的，因为使用“长”PCR方法，允许基因组DNA片段的定量扩增。

Kit Components

Component	Size / Description
2X Probe qPCR Master Mix	1 mL / 1 tube
Primer and Probe Mix	0.5 mL / 1 tube
Damaged DNA	10 µL/1 tube
Non-Damaged DNA	10 µL / 1 tube
Nuclease-free H2O	1 mL / 1 tube

Additional Materials Required

• Fluorescence PCR instrument capable of reading FAM channel (494 nm maximum absorption, 518 nm maximum emission) and JUN channel (606 nm maximum absorption, 618 nm maximum emission)
• Sterile nuclease-free pipette tips (barrier tips recommended) and microfuge tubes
• Compatible PCR Plate
• Microcentrifuge
• Vortex Mixer
• Pipettes

Assay Procedure Summary

1. Prepare all reagents, samples and standards as instructed.
2. Load the PCR reaction mix and samples.
3. Seal the PCR reaction tubes or plate.
4. Set up the PCR Amplification.
5. Export the results.
6. Calculate mtDNA copy number.

Specificity

The non-human and no DNA samples have higher Cq values than human DNA or are undetermined, which means there is high specificity for mtDNA amplification.

Reproducibility

CV <5% for Cq value and CV <15% for mtDNA damage when the input DNA >0.3 ng/reaction.

Typical Data

Storage/Stability

The kit can be stored at -20°C for a period of 12 months prior to opening. After opening, the reagents are valid for at least 6 months if stored at -20°C. Avoid repeated freeze-thaw cycles.