RayBio®线粒体DNA拷贝数绝对定量试剂盒（H-mtDNA-CNQ）是一种基于TaqmanTM探针的qPCR检测方法，用于人类mtDNA-CN的特定定量测量。 mtDNA引物和探针均特异性靶向人类mtDNA上最保守的区域之一，不会扩增出核基因组DNA上的任何脱靶序列。同时，新增单拷贝基因（IFNB1-人类 /36B4-小鼠）的引物和探针特异性识别和扩增110 bp长的核基因组DNA，作为数据归一化的参考。优化后的PCR配方在较宽的线性动态范围内提供了优越的特异性和效率。 本试剂盒可在一次反应中通过线粒体（mt）和核(n) DNA的比较来测定人类线粒体DNA拷贝数。

Kit Components

Component	Size / Description
2X Probe qPCR Master Mix	1 mL / 1 tube
Primer and Probe Mix	0.5 mL / 1 tube
Reference DNA (2500 copies/cell)	20 µL / 1 tube
Nuclease-free H2O	1 mL / 1 tube

Additional Materials Required

• Fluorescence qPCR instrument capable of reading FAM (494nm maximum absorption, 518nm maximum emission) and VIC (532nm maximum absorption, 552nm maximum excitation) channels
• Sterile nuclease-free pipette tips (barrier tips recommended)
• Microfuge tubes
• Compatible PCR Plate
• Microcentrifuge
• Vortex Mixer

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

Non-mouse DNA and no DNA samples have higher Cq values than mouse DNA, which means there is high specificity for mtDNA and 36B4 amplification.

Reproducibility

CV < 5% for Cq value and CV < 20% for CopiesCell when the input DNA ≥ 0.3 ngreaction.

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.