Evaluating the Efficiency of REPLI-g® Single Cell Kit for Trace DNA Amplification

doi:10.12116/j.issn.1004-5619.2019.02.015

科研成果详情

题名	Evaluating the Efficiency of REPLI-g® Single Cell Kit for Trace DNA Amplification
其他题名	Evaluating the Efficiency of REPLI-g~® Single Cell Kit for Trace DNA Amplification
作者	Q N Xu 1,2; Q Shen 3; J Y Zhang 2,4; Y L Zhang 2,5; L Li 2; X L Liu 2; C T Li 1,2
发表日期	2019-04-01
发表期刊	Fa yi xue za zhi 影响因子和分区
语种	英语
原始文献类型	Article
关键词	forensic genetics short tandem repeat whole genome amplification multiple displacement amplification trace samples.
摘要	Objective To evaluate the efficiency of REPLI-g® Single Cell Kit for sample DNA amplification, and explore its application value in forensic trace DNA amplification. Methods Three DNA extraction kits were selected to extract DNA from peripheral blood of 10 unrelated individuals. The DNA yield and purity of the three DNA extraction kits were compared. According to the results of comparison, one DNA sample was selected to concentrate and dilute, then used as the initial sample of whole genome amplification （WGA）. REPLI-g® Single Cell Kit was used to amplify the initial sample at the whole genome level. The amplification yield and amplification times were calculated, and the distribution of DNA fragments was detected by agarose gel electrophoresis. Goldeneye® DNA ID System 20A Kit was used to perform the STR typing of the initial sample and DNA samples amplified at the whole genome level to evaluate the performance of REPLI-g® Single Cell Kit in trace DNA amplication in terms of purity and yield as well as the success rate of STR typing. Results After comparison, one DNA sample was selected from QIAsymphony® DNA Investigator® Kit extracts to concentrate and dilute as the initial sample of WGA. After amplifying the whole genome of a series of initial samples by REPLI-g® Single Cell Kit, the lowest average of amplification yield reached 8.77×103 ng, while the average of the corresponding amplification times reached 1.40×106. DNA fragments were large and concentrated. The STR typing success rate of WGA samples became lower with the decrease of initial samples used, but when the amount of samples was lower than 0.5 ng, the STR typing success rate of samples after DNA WGA was higher than that of samples without DNA WGA. Conclusion REPLI-g® Single Cell Kit can increase the yield of template DNA. Especially for trace DNA, the STR typing success rate can be improved to a certain extent.
其他摘要	Objective To evaluate the efficiency of REPLI-g~® Single Cell Kit for sample DNA amplification, and explore its application value in forensic trace DNA amplification. Methods Three DNA extraction kits were selected to extract DNA from peripheral blood of 10 unrelated individuals. The DNA yield and purity of the three DNA extraction kits were compared. According to the results of comparison, one DNA sample was selected to concentrate and dilute, then used as the initial sample of whole genome amplification (WGA). REPLI-g ® Single Cell Kit was used to amplify the initial sample at the whole genome level. The amplification yield and amplification times were calculated, and the distribution of DNA fragments was detected by agarose gel electrophoresis. GoldeneyeTM DNA ID System 20A Kit was used to perform the STR typing of the initial sample and DNA samples amplified at the whole genome level to evaluate the performance of REPLI-g~® Single Cell Kit in trace DNA amplication in terms of purity and yield as well as the success rate of STR typing. Results After comparison, one DNA sample was selected from QIAsymphony® DNA Investigator® Kit extracts to concentrate and dilute as the initial sample of WGA. After amplifying the whole genome of a series of initial samples by REPLI-g ® Single Cell Kit, the lowest average of amplification yield reached 8.77×10~3 ng, while the average of the corresponding amplification times reached 1.40×10~6. DNA fragments were large and concentrated. The STR typing success rate of WGA samples became lower with the decrease of initial samples used, but when the amount of samples was lower than 0.5 ng, the STR typing success rate of samples after DNA WGA was higher than that of samples without DNA WGA. Conclusion REPLI-g~® Single Cell Kit can increase the yield of template DNA. Especially for trace DNA, the STR typing success rate can be improved to a certain extent.
资助项目	“十三五”国家重点研发计划资助项目 ; 国家自然科学基金 ; 中央级科研院所公益专项资助项目 ; 上海市标准研制资助项目 ; 上海市科委创新行动计划项目 ; 上海市法医学重点实验室资助项目 ; 上海市司法鉴定专业技术服务平台资助项目 ; 上海市人才项目
ISSN	1004-5619
卷号	35 期号:2 页码:210-215.
DOI	10.12116/j.issn.1004-5619.2019.02.015
页数	6
收录类别	PUBMED ; SCOPUS ; CSCD ; 万方 ; ISTIC
URL	查看原文
CSCD记录号	CSCD:6496845
PubMed ID	31135117
SCOPUSEID	2-s2.0-85066446748
引用统计
文献类型	期刊论文
条目标识符	https://kms.wmu.edu.cn/handle/3ETUA0LF/34002
专题	基础医学院（机能实验教学中心）_法医学系
作者单位	1.Department of Forensic Medicine, School of Basic Medical Sciences, Wenzhou Medical University, Wenzhou 325035, Zhejiang Province, China.; 2.Shanghai Key Laboratory of Forensic Medicine, Shanghai Forensic Service Platform, Academy of Forensic Science, Shanghai 200063, China.; 3.Department of Ultrasound Diagnosis, Shanghai Ninth People&s Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai 201999, China.; 4.Department of Forensic Medicine, Inner Mongolia Medical University, Hohhot 010030, China.; 5.Baotou Medical College, Inner Mongolia University of Science & Technology, Baotou 014060, Inner Mongolia Autonomous Region, China.
第一作者单位	基础医学院（机能实验教学中心）_法医学系
第一作者的第一单位	基础医学院（机能实验教学中心）_法医学系
推荐引用方式 GB/T 7714	Q N Xu,Q Shen,J Y Zhang,et al. Evaluating the Efficiency of REPLI-g® Single Cell Kit for Trace DNA Amplification[J]. Fa yi xue za zhi,2019,35(2):210-215..
APA	Q N Xu., Q Shen., J Y Zhang., Y L Zhang., L Li., ... & C T Li. (2019). Evaluating the Efficiency of REPLI-g® Single Cell Kit for Trace DNA Amplification. Fa yi xue za zhi, 35(2), 210-215..
MLA	Q N Xu,et al."Evaluating the Efficiency of REPLI-g® Single Cell Kit for Trace DNA Amplification".Fa yi xue za zhi 35.2(2019):210-215..

条目包含的文件		下载所有文件
文件名称/大小	文献类型	版本类型	开放类型	使用许可
FYXZ201902015.CAJ（1341KB）	期刊论文	出版稿	开放获取	CC BY-NC-SA	浏览下载