The paper aims to prepare the quality control for quantitative determination of residual DNA in E.coil.Extracting genomic DNA of Escherichia coli as a quality control material and analyzed for purity and concentration by UV spectrophotometry and agarose gel electrophoresis,the homogeneity and stability were checked by UV spectrophotometry and real-time PCR and evaluate its spiked recovery rate.The prepared quality control of Escherichia coli DNA was qualified as indicated by A260/A280 between 1.8 and 2.0 and a single specific band in agarose gel electrophoresis without RNA or oligonucleotides.The result showed the quality control materials were homogenous and stable under the-20 ℃ during 6 month's stock and 25 ℃ during 7 day's stock.The real-time PCR had high sensitivity up to 10-2-103 pg/μL of DNA with good linearity(r=0.997)with a slope of-3.65,and the recovery rate of each group of spiked samples was between 70％and 130％with the SRSD was less than or equal 20％.The prepared quality control was qualified in overall tests with DNA concentration was 111.73 µg/mL and may be used as quality control for residual DNA assay by real-time PCR methods.
万方数据
维普
