The Extraction of Metagenom DNA in Branch Bark

doi:10.13560/j.cnki.biotech.bull.1985.2016.01.018

Abstract

Abstract: A suitable method capable of extracting high-quality metagenom DNA from a variety of branch barks was developed based on previous experiences of plant DNA extraction. The method consisted of several steps：adding PVP while grinding materials, washing bark powder with bufferⅠ, simultaneously using SDS and CTAB, low-temperature frozen and then melting samples after adding high concentration of potassium acetate(KAc), precipitating DNA using isopropanol under high concentration of NaCl, and adding trace RNase A after DNA completely dissolved. Finally, the concentrations of genomic DNA obtained by this method ranged from 190 to 970 ng/μL with all in high purity. All DNA can be digested by restriction endonuclease and used as templates for bacterial 16S rRNA gene amplification. The DNA of apple bark as the template was amplified into gene BIP and PDI, then the genome DNA of apple bark was detected by a sequencing firm, and the quality satisfied the requirements of environmental metagenome for studying microbial diversity by high-throughput sequencing.

Key words: branch bark, DNA extraction, KAc, RNase A, high-throughput sequencing

ZHAO Ling-yun, FAN Dong-ying, LI Yan-fang, YAN Xia, HUANG Li-li. The Extraction of Metagenom DNA in Branch Bark[J]. Biotechnology Bulletin, 2016, 32(1): 74-79.

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