|本期目录/Table of Contents|

[1]陈振华,蔡甜,熊晓辉,等.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(02):102-108.[doi:10.3969/j.issn.1672-3678.2018.02.015]
 CHEN Zhenhua,CAI Tian,XIONG Xiaohui,et al.Dramatically enhancing direct detection from crude samples by Phanta DNA polymerase using optimized conditions[J].Chinese Journal of Bioprocess Engineering,2018,16(02):102-108.[doi:10.3969/j.issn.1672-3678.2018.02.015]
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优化反应体系提升Phanta DNA聚合酶对非处理标本的检测性能()
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《生物加工过程》[ISSN:1672-3678/CN:32-1706/Q]

卷:
16
期数:
2018年02期
页码:
102-108
栏目:
出版日期:
2018-03-30

文章信息/Info

Title:
Dramatically enhancing direct detection from crude samples by Phanta DNA polymerase using optimized conditions
文章编号:
1672-3678(2018)02-0102-07
作者:
陈振华1蔡甜1熊晓辉2许文玲3郭晓博3任顺利3韩锦雄3张海如34
1.佛山市南海区人民医院,广东 佛山 528000; 2.南京工业大学 食品与轻工学院,江苏 南京 211800; 3.南京诺唯赞生物科技有限公司,江苏 南京 210034; 4.武汉生物工程学院 生命科学与技术学院,湖北 武汉 430415
Author(s):
CHEN Zhenhua1CAI Tian1XIONG Xiaohui2XU Wenling3GUO Xiaobo3REN Shunli3HAN Jinxiong3ZHANG Hairu34
1.The People’s Hospital of Nanhai District,Foshan 528000,China; 2.College of Food Science and Light Industry, Nanjing Tech University,Nanjing 211800,China; 3.Nanjing Vazyme Biotechnology Co.,Ltd.,Nanjing 210034,China; 4.College of Biology and Biotechnology,Wuhan Institute of Bioengineering,Wuhan 430415,China
关键词:
Phanta DNA 聚合酶 PCR 添加剂 粗品 快速检测
分类号:
Q814.9
DOI:
10.3969/j.issn.1672-3678.2018.02.015
文献标志码:
A
摘要:
在食品检测、法医鉴定、分子诊断等领域经常需要对未提纯DNA的非规范处理标本进行聚合酶链式反应(PCR)扩增。然而,由于非处理标本中存在多种抑制成分,常见的DNA聚合酶并不能满足非处理标本的检测需求。本研究通过调节添加剂、Mg2+浓度以及酶量,对Phanta DNA聚合酶进行反应条件优化,提升Phanta DNA聚合酶突变体的扩增性能,并考察该聚合酶在优化条件下对血液、植物和食品等非处理标本的检测性能。结果表明:在反应缓冲溶液SF Buffer中加入质量分数5%二甲基亚砜(DMSO)和3 mmol/L Mg2+,以及50 μL反应体系中使用1 U酶时,Phanta DNA聚合酶突变体具有最强的扩增性能,可对低至0.05 ng基因组、植物叶片粗提液、高达40%全血浓度及食品粗提液进行有效检测,性能优于普通Taq DNA聚合酶及未进行反应条件优化的Phanta DNA聚合酶。本研究在食品检测、法医鉴定、分子诊断等应用中非处理标本的检测具有极大应用价值。

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备注/Memo

备注/Memo:
收稿日期:2018-02-28修回日期:2018-03-11
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)
作者简介:陈振华(1971—),男,广东佛山人,副主任医师,研究方向:分子检测、呼吸内科,E-mail:282090822@qq.com
引文格式:陈振华,蔡甜,熊晓辉.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.
CHEN Zhenhua,CAI Tian,XIONG Xiaohui,et al.Dramatically enhancing direct detection from crude samples by Phanta DNA polymerase using optimized conditions[J].Chin J Bioprocess Eng,2018,16(2):102-108..
更新日期/Last Update: 2018-03-30