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- 详细信息
- 技术资料
- 英文名：
Human Mitochondrial DNA Damage Quantification qPCR Assay Kit
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
- Prepare all reagents, samples and standards as instructed.
- Load the PCR reaction mix and samples.
- Seal the PCR reaction tubes or plate.
- Set up the PCR Amplification.
- Export the results.
- 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.
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人类线粒体DNA损伤 qPCR试剂盒
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