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| Isothermal detection of RNA transcription levels using graphene oxide-based SYBR Green I fluorescence platform |
| Wen-Ge Maa, Su-Qin Lub, De-Bin Zhuc, Xiao-Bo Xingc, Bi-Hua Sud |
a Guangzhou Nansha Center for Disease Control & Prevention, Guangzhou 511457, China;
b Department of Medical Ultrasonics, The Third Affiliated Hospital, Sun Yat-Sen University, Guangzhou 510630, China;
c MOE Key Laboratory of Laser Life Science & Institute of Laser Life Science, College of Biophotonics, South China Normal University, Guangzhou 510631, China;
d Guangzhou Baiyun District Center for Disease Control & Prevention, Guangzhou 510445, China |
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Abstract An isothermal graphene oxide (GO)-based SYBR Green I fluorescence platform for sensitive detection of RNA transcription levels is proposed. Briefly, a synthesized T7 DNA template was transcribed by T7 RNA polymerase, to producemany copies of single-stranded (ss) RNA transcripts. The ssRNA transcripts were then hybridized with a label-free ssDNA probe which alone will be adsorbed on the GO surface. The resultant double-stranded (ds) RNA:DNA hybrids were incubated with GO and SYBR Green I. Then the hybrid binding with SYBR Green I emits enhanced fluorescence for detection. Experimental results show that the detection limit of the method is 0.5 pmol/L of T7 DNA template. A calibration curve with a linearity range of 0.5 pmol/L to 5 nmol/L is established, thus, making quantitative analysis feasible. The method may become a powerful tool for RNA transcription detection due to its sensitivity, rapidity and convenience.
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Received: 04 November 2013
Published: 24 December 2013
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| Fund: This research is supported by the National Natural Science Foundation of China (Nos. 81071790, 81371877, 61177077), the Key Project of Chinese Ministry of Education (No. 211131), the Science and Technology Project of Guangzhou Nansha District (No. RG201001003), and the Postdoctoral Foundation of China (No. 201003359). |
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Corresponding Authors:
Wen-Ge Ma
E-mail: nsmwg@163.com
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