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An Improved Protocol for Low Copy Number Plasmid DNA Purification with Diatomaceous Earth
Wen-Jie Tang1, 2, Ma-Lin Li1, Chui-Yuan Qiu2, Qiong-Yu Chen2, Guo-Hui Li, Ling-Song Li3, An Hong2*
1Yunnan Pharmacological Laboratory of Natural Products, Kunming Medical College, Kunming 650031, China;2Bioengineeering Institute of Jinan University, Guangzhou 510632, China;3The Centre of Stem Cell Research, University o
Abstract: Sequencing of low copy number plasmid DNA failed quite often due to high amount of impurities in sample. To develop an efficient and highly repeatable DNA purification protocol especially suitable for low copy number plasmid DNA， the amount of diatomaceous earth suspension applied to bind DNA was optimized. It was found that 1 mg of pure lDNA was most efficiently recovered when 20 ml of the suspension was applied， with a recovery as high as 77.31%. pLZ14， a low copy number plasmid of 14 953 bp， was purified 12.06-fold with the improved protocol， reaching a purity of 23.87%. Strong， clear sequencing signals and correct results were obtained with the purified pLZ14 plasmid DNA， while weak， confusing signals and universal errors were found for DNA from routine alkaline lysis and purification protocols.
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