Team:XHD-WS-Wuhan-A/Model
Model
The effect of miR-196a-148a sensor as a monitor to detect the expression of miR-148a
To test the effect of miR-196a-148a sensor as a monitor to detect the expression of miR-148a, cells were transfected with pEGFP-miR-196a-148a sensor with different concentration of pHAGE-pre-miR-148a. We used the plasmid of pHAGE-pre-miR-148a to quantify miR-148a expression and calculate the copy numbers of miR-148a by using the formula listed below.
copies/ul= (6.02×1023)×(plasmids concentrations ng/ul×10-9)/(DNA length×660)
Table 3 The value of eGFP fluorescence
|
Copies/ul |
1.755×107 |
2.34×107 |
2.925×107 |
3.51×107 |
|
GFP |
1141899 |
1007097 |
944104 |
903925 |
|
1113005 |
1022613 |
891213 |
840968 |
|
|
1199842 |
1035901 |
978254 |
873048 |
|
|
1129558 |
1071912 |
951016 |
784177 |
|
|
AVERAGE |
1146076 |
1034380.75 |
941146.75 |
850529.5 |
The standard curve of miR-196a-148a sensor was made by EXCEL (Fig 17). We find that the value of fluorescence is dependent on the copy numbers of miR-148a in cells. Based on the formula, the correlation coefficient (R2 value) of miR-196a-148a sensor was 0.9974.
Fig 17. The standard curve of miR-196a-148a sensor
References:
1. Zheng G, Xiong Y, Xu W, Wang Y, Chen F, Wang Z, Yan Z. A two-microRNA signature as a potential biomarker for early gastric cancer. Oncol Lett. 2014 Mar;7(3):679-684.
2. Song JH, Meltzer SJ. MicroRNAs in pathogenesis, diagnosis, and treatment of gastroesophageal cancers. Gastroenterology. 2012 Jul;143(1):35-47.
3. Sierzega M, Kaczor M, Kolodziejczyk P, Kulig J, Sanak M, Richter P. Evaluation of serum microRNA biomarkers for gastric cancer based on blood and tissue pools profiling: the importance of miR-21 and miR-331. Br J Cancer. 2017 Jul 11;117(2):266-273.
Address
XHD-WS-Wuhan-A
Wuhan High School
No.538,Gongxiu Road,Xiaoshan District,
Wuhan,Hubei Province,China