Pyrosequencing Detection Pass Ratio for DNA Methylation

Zhao Fei; Jiang Yan Ling; Zhang Shou Xun; Xia Sheng; Bao Jian Jun; Zhang Nin; Yang Xiao Pei; Zhong Shu Rong

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Article Navigation > Journal of Kunming Medical University > 2017 > 38(10): 27-31

Zhao Fei , Jiang Yan Ling , Zhang Shou Xun , Xia Sheng , Bao Jian Jun , Zhang Nin , Yang Xiao Pei , Zhong Shu Rong . Pyrosequencing Detection Pass Ratio for DNA Methylation[J]. Journal of Kunming Medical University, 2017, 38(10): 27-31.

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Zhao Fei , Jiang Yan Ling , Zhang Shou Xun , Xia Sheng , Bao Jian Jun , Zhang Nin , Yang Xiao Pei , Zhong Shu Rong . Pyrosequencing Detection Pass Ratio for DNA Methylation[J]. Journal of Kunming Medical University, 2017, 38(10): 27-31.

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Pyrosequencing Detection Pass Ratio for DNA Methylation

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基金：国家自然科学基金资助项目 (81660232, 81000577); 云南省科技厅-昆明医科大学应用基础研究联合专项基金资助项目 (2015FB011); 昆明医科大学大学生创新性试验计划项目 (CX201416, CX201654, 601162023);

Received Date: 2017-05-21

Abstract

Abstract

Objecctive To analyze the result of the pass ratio when we use different dose (2.5 μL, 5 μL and 10 μL) of PCR products to detect DNA methylation by pyrosequencing. Methods DNA extracted from blood samples that collected from 20 alcohol-dependent patients and 20 healthy controls were treated with Epi Tect Fast DNA Bisulfite Kit. PCR amplification was performed using converted DNA as template.We added different dose of PCR products (2.5 μL, 5 μL, 10 μL) to detecte DNA methylation quantification values of target gene. Results By comparing the average peak height and the pass ratio of pyrosequencing, we found the group (10 μL) was the maximum, the group (2.5 μL) was the maximum. The result of the average peak height and the pass ratio of three groups was increased significantly (P<0.05) . Conclusions When we treat 200 ng DNA extracted from blood samples with Epi Tect Fast DNA Bisulfite Kit, the group (10 μL) sequencing pass ratio is optimum. The sequencing pass ratio of the group (10 μL) is 100 %.
- Pyrosequencing,
- Detection pass ratio,
- DNA Methylation

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References(20)

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