Comparison of Two Extraction Methods in Extraction of Total DNA from Soil Microbes in the Rhizosphere of Pitaya

CHEN Di; LI Kezeng; WU Qiong; HE Yindui; YIN Xiaomin

doi:10.15886/j.cnki.rdswxb.2019.02.014

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In order to obtain the total DNA of the soil microbes in the rhizosphere of pitaya（Hylocereus undatus Britt.）, two methods, CTAB-SDS and SDS, were used to compare their extraction effect in extraction time, color of DNA supernatant in the extraction process, and total DNA extraction amount and purity after extraction. The results showed that both methods produced DNA fragments with about 23.1 kb. With the CTAB-SDS method the color of the supernatant was light yellow; the total amount of DNA extracted was（7.4 ～ 7.7） mg·kg^-1; the OD_260/280 value was between 1.81 and 1.95, and the OD_260/230 value was between 1.26 and 2.03; the average DNA concentration was 0.51 g·L^-1. With the SDS method the color of the supernatant was dark brown-yellow, and the total amount of DNA extracted was（3.5 ～ 4.15） mg·kg^-1; the OD_260/280 value was between 1.52 and 1.68, and the OD_260/230 value was between 1.35 and 1.53; the average DNA concentration was 0.075 g·L^-1. It was hence concluded that the fragment extracted by the CTAB-SDS method was relatively complete and highly pure, which could be directly used for subsequent experimental operation, and was more suitable for the extraction of total DNA from soil microbes in the rhyzosphere of pitaya.

Comparison of Two Extraction Methods in Extraction of Total DNA from Soil Microbes in the Rhizosphere of Pitaya

1. Haikou Experimental Station, CATAS, Haikou, Hainan 570102, China;

2. Hainan Key Laboratory of Banana Genetic Improvement, CATAS, Haikou, Hainan 570102 China;

3. Institute of Tropical Agriculture and Forestry, Hainan University, Haikou, Hainan 570102, China

Received Date: 2018-11-27

Rev Recd Date: 2019-04-08

Key words:

Abstract: In order to obtain the total DNA of the soil microbes in the rhizosphere of pitaya（Hylocereus undatus Britt.）, two methods, CTAB-SDS and SDS, were used to compare their extraction effect in extraction time, color of DNA supernatant in the extraction process, and total DNA extraction amount and purity after extraction. The results showed that both methods produced DNA fragments with about 23.1 kb. With the CTAB-SDS method the color of the supernatant was light yellow; the total amount of DNA extracted was（7.4 ～ 7.7） mg·kg^-1; the OD_260/280 value was between 1.81 and 1.95, and the OD_260/230 value was between 1.26 and 2.03; the average DNA concentration was 0.51 g·L^-1. With the SDS method the color of the supernatant was dark brown-yellow, and the total amount of DNA extracted was（3.5 ～ 4.15） mg·kg^-1; the OD_260/280 value was between 1.52 and 1.68, and the OD_260/230 value was between 1.35 and 1.53; the average DNA concentration was 0.075 g·L^-1. It was hence concluded that the fragment extracted by the CTAB-SDS method was relatively complete and highly pure, which could be directly used for subsequent experimental operation, and was more suitable for the extraction of total DNA from soil microbes in the rhyzosphere of pitaya.

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Reference (18)

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Comparison of Two Extraction Methods in Extraction of Total DNA from Soil Microbes in the Rhizosphere of Pitaya

doi: 10.15886/j.cnki.rdswxb.2019.02.014

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