一种适用于高通量测序的红曲霉基因组DNA提取方法

Optimized method for the preparation of Monascus purpureus genomic DNA for high-throughput sequencing

通过比较5种红曲霉基因组DNA的提取方法,旨在获得一种简便高效、大量提取高质量基因组DNA的优化方法。分别使用改进CTAB法、改进氯化苄法、蜗牛酶法、SDS法、试剂盒法提取红曲霉基因组DNA,结果显示:除蜗牛酶外,其余4种方法均能够提取出红曲霉基因组DNA,其中利用改进CTAB法能够快速、经济地得到大量高纯度的DNA样品,完全满足高通量测序、分子标记等一系列精密分子生物学研究的要求。

In order to find an optimized and effective method, there were five methods of Monascus purpureus genomic DNA extraction to be compared, including CTAB protocol with some modifications, benzyl chloride protocol with some modifications, snailase protocol, SDS protocol and extraction with the E.Z.N.A. Fungal DNA Maxi Kit. Results demonstrated that genomic DNA of Monascus purpureus could be isolated by the four methods except snailase protocol. The CTAB method which we established with some modification in several steps was not only economical and convenient, but also could be reliably used to obtain large amounts of highly pure genomic DNA from Monascus purpureus for precise molecular biology researches such as high-throughput sequencing and molecular marker.