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  本文選題：腸道菌群 + 鳥(niǎo)槍法測(cè)序�。� 參考：《中國(guó)生物工程雜志》2017年11期

【摘要】：目的:優(yōu)化腸道菌群核酸提取自動(dòng)化流程。方法:收集糞便樣本,分別采用手工方法、核酸提取工作站提取核酸,然后利用液體工作站制備PCR反應(yīng)液進(jìn)行PCR擴(kuò)增,最后采用文庫(kù)工作站建庫(kù)測(cè)序。結(jié)果:400μl樣品用QIAamp~汶 Fast DNA Stool Mini Kit試劑盒裂解液裂解后,用MagMAX~(TM) Express 96機(jī)器提取核酸的方法與用QIAamp~汶 Fast DNA Stool Mini Kit試劑盒手工提取核酸的方法相比,測(cè)序得到的序列數(shù)基本一致,分析結(jié)果也沒(méi)有明顯區(qū)別;而單獨(dú)采用MagMAX~(TM)Viral RNA Isolation Kit試劑盒提取核酸由于樣品投入體積受限(50μl)、核酸濃度低、測(cè)序得到的序列數(shù)太少,不能滿(mǎn)足后續(xù)的分析要求。結(jié)論:通過(guò)結(jié)合使用兩種不同的試劑盒,可以實(shí)現(xiàn)核酸提取、PCR反應(yīng)液配制及文庫(kù)制備的全流程自動(dòng)化操作,從而大大提高工作效率和結(jié)果的穩(wěn)定性。
[Abstract]:Objective: to optimize the automated extraction process of intestinal bacterial nucleic acid. Methods: the fecal samples were collected, the nucleic acid was extracted by manual method, and the PCR reaction solution was prepared by liquid workstation for PCR amplification. Finally, the library was constructed and sequenced by library workstation. Results the nucleic acid was extracted by using MagMAXT Express 96 machine after the cleavage of 1: 400 渭 l sample with QIAamp-Wen Fast DNA Stool Mini Kit kit. Compared with the method of manual extraction of nucleic acid with QIAamp-Fast DNA Stool Mini Kit kit, the sequence number obtained by sequencing was basically the same. There was no significant difference in the analysis results, but the MagMAX~(TM)Viral RNA Isolation Kit kit alone was used to extract nucleic acid because of the limited input volume of 50 渭 l of nucleic acid, the low concentration of nucleic acid, and the small number of sequencing sequences, which could not meet the requirements of subsequent analysis. Conclusion: by combining two different kits, the whole process of preparation of nucleic acid extraction PCR reaction solution and the preparation of library can be realized, thus greatly improving the working efficiency and stability of the results.
【作者單位】： 安徽醫(yī)科大學(xué);中國(guó)人民解放軍軍事科學(xué)院微生物與流行病研究所;
【基金】：病原微生物生物安全國(guó)家重點(diǎn)實(shí)驗(yàn)室課題資助項(xiàng)目(SKLPBS1518)
【分類(lèi)號(hào)】：R37

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本文編號(hào)：1882142