生物加工过程2018，Vol.16Issue(2)：102-108,7.DOI:10.3969/j.issn.1672-3678.2018.02.015
优化反应体系提升Phanta DNA聚合酶对非处理标本的检测性能
Dramatically enhancing direct detection from crude samples by Phanta DNA polymerase using optimized conditions
摘要
Abstract
Polymerase chain reaction were applied in food detection,forensic identification,molecular diagnosis,and so on.In these applications,lots of samples were crude samples without DNA purification. These samples had many inhibitors which would inhibit PCR reaction. To enhance direct DNA amplification from crude samples,like blood,plant,and food,reaction condition of Phanta DNA polymerase was optimized by PCR additives,gradient concentration and polymerase content.DMSO and magnesium were used as PCR additives and the optimal concentration was 5% DMSO and 3 mmol/L magnesium ion.The optimized reaction condition,in combination with 1 U Phanta DNA polymerase in 50 μL reaction volum,enabled efficient detection of PCR products from as low as 0.050 ng human genome DNA,40% blood,leaf,and food crude extracts.Moreover,compared with commercial DNA polymerases, the novel PCR additives coordinate with Phanta DNA polymerase had better performance.This study has important application value of crude sample detection in food detection,forensic identification and molecular diagnosis.关键词
PhantaDNA聚合酶/PCR/添加剂/粗品/快速检测Key words
Phanta DNA polymerase/PCR/additives/crude samples/rapid detection分类
生物科学引用本文复制引用
陈振华,蔡甜,熊晓辉,许文玲,郭晓博,任顺利,韩锦雄,张海如..优化反应体系提升Phanta DNA聚合酶对非处理标本的检测性能[J].生物加工过程,2018,16(2):102-108,7.基金项目
江苏省重点领域共性技术攻关项目(苏经信科技(2016)636号) （苏经信科技(2016）
