Analysis of the inhibition of nucleic acid dyes on polymerase chain reaction by capillary electrophoresis

被引:6
|
作者
Li, Zhenqing [1 ]
Liu, Chenchen [1 ]
Ma, Siyao [1 ]
Zhang, Dawei [1 ]
Yamaguchi, Yoshinori [2 ,3 ]
机构
[1] Univ Shanghai Sci & Technol, Engn Res Ctr Opt Instrument & Syst, 516 Jungong Rd, Shanghai 200093, Peoples R China
[2] ECUST, IPBM, Grad Sch Sci, 130 Meilong Rd, Shanghai 200237, Peoples R China
[3] Osaka Univ, Dept Appl Phys, Grad Sch Engn, 2-2 Yamadaoka, Suita, Osaka 5650871, Japan
基金
高等学校博士学科点专项科研基金; 中国国家自然科学基金;
关键词
SYBR-GREEN-I; REAL-TIME PCR; PORPHYROMONAS-GINGIVALIS; ENZYMATIC AMPLIFICATION; PERIODONTAL PATHOGENS; ESCHERICHIA-COLI; DNA; ASSAY; VIRUS; QUANTIFICATION;
D O I
10.1039/c5ay02705e
中图分类号
O65 [分析化学];
学科分类号
070302 ; 081704 ;
摘要
An integrated polymerase chain reaction (PCR) and capillary electrophoresis (CE) system can realize accurate quantification of the target PCR product by adding labeling dyes to the PCR reagents, because CE can discriminate all the subsequent nucleic acids, including the primers, non-specific and specific PCR products. Here we discuss the inhibition of labeling dyes on PCR by performing the PCR of Porphyromonas gingivalis (PG) with solutions containing Hoechst 33258, SYBR Green I, and SYBR Green II. Results demonstrated that Hoechst 33258 totally inhibited the PCR process, and PCR efficiency was highly dependent on the concentration of SYBR Green I/II. Such a study expands the capabilities of CE and contributes greatly to the development of hyphenated PCR-CE instruments for biological and medical diagnosis.
引用
收藏
页码:2330 / 2334
页数:5
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