Isothermal detection of RNA transcription levels using graphene oxide-based SYBR Green I fluorescence platform

Citation: Wen-Ge Ma, Su-Qin Lu, De-Bin Zhu, Xiao-Bo Xing, Bi-Hua Su. Isothermal detection of RNA transcription levels using graphene oxide-based SYBR Green I fluorescence platform[J]. Chinese Chemical Letters, 2014, 25(3): 482-484. doi: 10.1016/j.cclet.2013.12.013 shu

Isothermal detection of RNA transcription levels using graphene oxide-based SYBR Green I fluorescence platform

通讯作者: Wen-Ge Ma,

基金项目:
This research is supported by the National Natural Science Foundation of China (Nos. 81071790, 81371877, 61177077) (Nos. 81071790, 81371877, 61177077)

the Key Project of Chinese Ministry of Education (No. 211131) (No. 211131)

Technology Project of Guangzhou Nansha District (No. RG201001003) (No. RG201001003)

the Postdoctoral Foundation of China (No. 201003359). (No. 201003359)

摘要: 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.

关键词:

English

Isothermal detection of RNA transcription levels using graphene oxide-based SYBR Green I fluorescence platform

1.
a Guangzhou Nansha Center for Disease Control & Prevention, Guangzhou 511457, China;
2.
b Department of Medical Ultrasonics, The Third Affiliated Hospital, Sun Yat-Sen University, Guangzhou 510630, China;
3.
c MOE Key Laboratory of Laser Life Science & Institute of Laser Life Science, College of Biophotonics, South China Normal University, Guangzhou 510631, China;
4.
d Guangzhou Baiyun District Center for Disease Control & Prevention, Guangzhou 510445, China

Corresponding author: Wen-Ge Ma,

Received Date: 04 November 2013
Available Online: 24 December 2013
Fund Project:

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.

Key words:

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沈阳化工大学材料科学与工程学院沈阳 110142

Isothermal detection of RNA transcription levels using graphene oxide-based SYBR Green I fluorescence platform

通讯作者: Wen-Ge Ma,

English

Isothermal detection of RNA transcription levels using graphene oxide-based SYBR Green I fluorescence platform

Corresponding author: Wen-Ge Ma,

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通讯作者: 陈斌, bchen63@163.com

中国化学会秘书处

Isothermal detection of RNA transcription levels using graphene oxide-based SYBR Green I fluorescence platform

通讯作者: Wen-Ge Ma,

English

Isothermal detection of RNA transcription levels using graphene oxide-based SYBR Green I fluorescence platform

Corresponding author: Wen-Ge Ma,

CCS Chemistry：嵌段共聚物自组装成 “三明治”二维有机材料，实现纳米级压力传感功能

CCS Chemistry：颜徐州、鲍哲南： 你的皮肤可能是一片SPMs

CCS Chemistry：工作高效不疲劳，只须让催化剂动起来

CCS Chemistry：静电组装导向聚合- 聚电解质纳米凝胶量化制备新策略

CCS Chemistry：金黄色葡萄球菌醛基脱氢酶是如何识别特异性底物的？

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