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| Optimization of Automatic Flow Chart for Nucleic Acid Extraction from Intestinal Microflora |
| LI Jin-man1,2, PEI Guang-qian2, FAN Hang2, HUANG Yong2, TONG Yi-gang1,2 |
1. Anhui Medical University, Hefei 230032, China;
2. Institute of Microbiology and Epidemiology, the Academy of Military Sciences PLA China, Beijing 100071, China |
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Abstract Objective:Optimize the automatic flow of nucleic acid extraction from intestinal flora. Methods:Fecal samples were collected, and nucleic acids were extracted by manual method and nucleic acid extraction workstation. Then PCR reaction liquid was prepared by liquid workstation, and PCR was amplified and sequenced by library workstation. Results:400μl samples using QIAamp® Fast DNA Stool Mini Kit, compared with nucleic acid extraction method with QIAamp® Fast DNA Stool Mini Kit manual with MagMAXTM Express 96 machine nucleic acid extraction method, sequence number sequencing analysis results obtained are basically the same, there were also no significant differences; the single use of MagMAXTMViral RNA Isolation Kit to extract nucleic acid sequencing results due to sample input limited volume (50μl), low concentration of nucleic acid and littlesequence number, can't meet the requirements of the follow-up analysis.Conclusion:By using two different kits, the whole process automation of nucleic acid extraction, PCR reaction preparation and library preparation can be realized, thus greatly improving the efficiency and stability of the results.
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Received: 20 September 2017
Published: 15 November 2017
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