A novel assay coupling dephosphorylation and blue/white colony screening for the G >a hotspot mutation at codon 13 of KRAS gene

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成果类型：

期刊论文

作者：

Zhou, Cuilan;Pu, Wangyang;Yin, Yu-Fang;Xiao, Li;Luo, Dixian;...

通讯作者：

Cai, Weijun;Liao, Duanfang

作者机构：

[Zhou, Cuilan; Cai, Weijun; Liao, DF; Liao, Duanfang] Cent S Univ, Sch Basic Med, Dept Histol & Embryol, Changsha 410078, Hunan, Peoples R China.

[Zhou, Cuilan; Guo, Zifen; Peng, Cuiying] Univ South China, Dept Human Anat, Hengyang 421001, Peoples R China.

[Yin, Yu-Fang] Soochow Univ, Coll Pharmaceut Sci, Dept Mol Diagnost & Biopharmaceut, Suzhou 215123, Peoples R China.

[Li, Kai; Xing, Chungen; Sun, Wanping; Pu, Wangyang; Liu, Xuan; Wang, Fenjiao; Xu, Huifen; Xiao, Li] Univ South China, Peoples Hosp Chenzhou 1, Translat Med Inst, Natl & Local Joint Engn Lab High Throughput Mol D, Hengyang 423000, Peoples R China.

[Luo, Dixian; He, Nongyue] Southeast Univ, Sch Biol Sci & Med Engn, State Key Lab Bioelect, Nanjing 211189, Jiangsu, Peoples R China.

通讯机构：

[Cai, WJ; Liao, DF] C

Cent S Univ, Sch Basic Med, Dept Histol & Embryol, Changsha 410078, Hunan, Peoples R China.

语种：

英文

关键词：

KRAS;Sanger's Sequencing;Blue/White Screening;ctDNA;Cancer Test

期刊：

JOURNAL OF NANOSCIENCE AND NANOTECHNOLOGY

ISSN：

1533-4880

年：

2018

卷：

期：

页码：

538-543

DOI：

10.1166/jnn.2018.13935

基金类别：

National Natural Science Foundation of ChinaNational Natural Science Foundation of China (NSFC) [81370248]; Priority Academic Program Development of Jiangsu Higher Education Institutions (PAPD); Jiangsu Province's clinical medical science and technology program [BL2013016]; natural science foundation of Hunan ProvinceNatural Science Foundation of Hunan Province [2016JJ6126, 2017JJ3280]

机构署名：

本校为其他机构

院系归属：

医学院

摘要：

Development of sensitive assay for detection of hotspot mutations of cancer driving gene is crucial for circulating tumor DNA analysis. This study tested the possibilities of applying restriction enzyme digestion and dephosphorylation coupled with blue/white screening technology for analyzing a hotspot point mutation in codon 13 of KRAS gene. The present study has documented that the combination of PCR with restriction digestion, dephosphorylation, blue/white screening and Sanger's sequencing can identify rare mutations with sensitivities at 0.003%. This novel assay with high sensitivity may h...

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A novel assay coupling dephosphorylation and blue/white colony screening for the G >a hotspot mutation at codon 13 of KRAS gene

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