不同扩增方法对豆浆微生物PCR-DGGE检测的影响
Effects of the Amplification Methods on the Detection of Microorganisms in Soymilks by PCR-DGGE
大豆中的叶绿体16S rRNA基因会干扰微生物16S rDNA片段的扩增。为消除叶绿体的干扰，提高微生物检测限，本实验选择四种不同引物对豆浆DNA进行扩增：1.使用引物F341-GC/R534直接扩增V3区；2. 使用引物27F/1492R先扩增16S全长，再使用引物F341-GC/R534扩增V3区；3. 使用引物27F/783R扩增V2-V3区，再使用引物F341-GC/R534扩增V3区；4. 使用引物HDA4-GC/HDA5扩增V2区。通过PCR-DGGE检测叶绿体的去除情况，微生物的丰富度及对于豆浆中优势菌的检测限，结果表明方法3为最佳扩增方法。
In order to eliminate the interference of chloroplast and improve the detection limit of microorganism, four DNA amplification methods were selected in this experiment: 1. Using primer f341-gc / r534 to amplify V3 region directly. 2. Primer 27F / 1492r was used to amplify the whole length of 16S, and then V3 region. 3. The v2-v3 region was amplified by primer 27F / 783r, and V3 region was amplified by primer f341-gc / r534. 4. Using primer hda4-gc / hda5 to amplify V6 region. PCR-DGGE was used to detect the removal of chloroplast, the abundance of microorganisms and the detection limit of dominant bacteria in soybean milk. The results showed that method 3 was the best amplification method.
尹梦琪、华欲飞、孔祥珍、张彩猛、李兴飞、陈业明
微生物学生物科学研究方法、生物科学研究技术分子生物学
叶绿体引物选择PCR-DGGE检测限
hloroplastPrimer selectionPCR-DGGEetection limit
尹梦琪,华欲飞,孔祥珍,张彩猛,李兴飞,陈业明.不同扩增方法对豆浆微生物PCR-DGGE检测的影响[EB/OL].(2020-04-28)[2025-08-16].http://www.paper.edu.cn/releasepaper/content/202004-289.点此复制
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