材料与试剂

1. 移液器吸头、离心管
   
2. 96孔板和封板膜 (Roche, LightCycler 480 Multiwell Plate 96)
   
3. DNA提取试剂盒 (MP Biomedicals, FastDNA Spin Kit)
   
4. 高保真PCR酶试剂 (New England Biolabs, Phusion High-Fidelity PCR Master Mix)
   
5. 电泳上样缓冲液Loading buffer (Takara, 6× Loading Buffer, Cat. No. 9156)
   
6. 琼脂糖粉末 (Sigma, A9539-250G)
   
7. TAE溶液 (Solarbio, 50× TAE)
   
8. DNA Marker (New England Biolabs, 50 bp DNA Ladder, NEB #B7025)
   
9. 凝胶纯化试剂盒 (南京诺唯赞, DC301-01)
   
10. Qubit分子定量试剂盒 (Promega, QuantiFlour dsDNA System)
    

仪器设备

1. 移液器 (Eppendorf, 100–1000 μl, 10–100 μl 20–200 μl, 2–20 μl, 0.5–10 μl, 0.1–2.5 μl)
   
2. 均质仪 (杭州奥盛, Bioprep-24)
   
3. 离心机 (Eppendorf, 5417R)
   
4. 干式加热器 (Labnet, D1100-230V)
   
5. 超净工作台 (苏净安泰, Airtech)
   
6. 漩涡混合器 (海门其林贝尔, Vortex-6)
   
7. 200 μl八联移液器 (Brand, 10-200 μl)
   
8. 96孔板离心机 (杭州奥盛, Mini-P25)
   
9. PCR热循环仪 (Bio-Rad, S1000)
   
10. 电泳仪 (北京六一, DYY6C)
    
11. 电泳槽 (北京六一, JYCP31DN)
    
12. 凝胶成像仪 (上海培清, JS-680B)
    
13. 核酸测定仪 (ThermoFisher, Qubit 4.0)
    

实验步骤