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比较从珊瑚礁中提取微生物 DNA 的不同方案。

Comparison of different protocols for the extraction of microbial DNA from reef corals.

机构信息

Laboratório de Ecologia Molecular Microbiana, Universidade Federal do Rio de Janeiro , Rio de Janeiro, RJ , Brasil.

出版信息

Braz J Microbiol. 2012 Apr;43(2):517-27. doi: 10.1590/S1517-83822012000200012. Epub 2012 Jun 1.

Abstract

This study aimed to test different protocols for the extraction of microbial DNA from the coral Mussismilia harttii. Four different commercial kits were tested, three of them based on methods for DNA extraction from soil (FastDNA SPIN Kit for soil, MP Bio, PowerSoil DNA Isolation Kit, MoBio, and ZR Soil Microbe DNA Kit, Zymo Research) and one kit for DNA extraction from plants (UltraClean Plant DNA Isolation Kit, MoBio). Five polyps of the same colony of M. harttii were macerated and aliquots were submitted to DNA extraction by the different kits. After extraction, the DNA was quantified and PCR-DGGE was used to study the molecular fingerprint of Bacteria and Eukarya. Among the four kits tested, the ZR Soil Microbe DNA Kit was the most efficient with respect to the amount of DNA extracted, yielding about three times more DNA than the other kits. Also, we observed a higher number and intensities of DGGE bands for both Bacteria and Eukarya with the same kit. Considering these results, we suggested that the ZR Soil Microbe DNA Kit is the best adapted for the study of the microbial communities of corals.

摘要

本研究旨在测试从珊瑚 Mussismilia harttii 中提取微生物 DNA 的不同方案。测试了四种不同的商业试剂盒,其中三种基于从土壤中提取 DNA 的方法(FastDNA SPIN 试剂盒用于土壤,MP Bio,PowerSoil DNA 分离试剂盒,MoBio 和 ZR 土壤微生物 DNA 试剂盒,Zymo Research),一种试剂盒用于从植物中提取 DNA(UltraClean 植物 DNA 分离试剂盒,MoBio)。将同一珊瑚礁 M. harttii 同一殖民地的五个息肉捣碎,并将等分试样提交给不同试剂盒进行 DNA 提取。提取后,对 DNA 进行定量,并使用 PCR-DGGE 研究细菌和真核生物的分子指纹。在测试的四个试剂盒中,ZR 土壤微生物 DNA 试剂盒在提取的 DNA 量方面最有效,其产量比其他试剂盒高约三倍。同样,我们还观察到相同试剂盒的细菌和真核生物的 DGGE 带的数量和强度更高。考虑到这些结果,我们建议 ZR 土壤微生物 DNA 试剂盒最适合研究珊瑚的微生物群落。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/bb0f/3768815/702c1fee2630/bjm-43-517-g001.jpg

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