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Shen Yangli,Sun Hao,Cai Wenchen,Zhang Pinzheng,Wang Xuying,Shang Yuhan,Shi Lu,Guo Zhiyi.Assessment of quality of tri-allelic polymorphism compassing rs6983267 based on droplet digital PCR taking advantage of probe cross-reactivity<sub>*</sub>[J].Progress in Biochemistry and Biophysics
Assessment of quality of tri-allelic polymorphism compassing rs6983267 based on droplet digital PCR taking advantage of probe cross-reactivity<sub>*</sub>
Received:July 09, 2018  Revised:August 16, 2018
Key words:digital PCR  nucleotide acid polymorphism  cross-reactivity probe  rs6983267
Fund:国家自然科学基金(编号:31050010),教育部归国留学基金(编号:[2015]No.311),华北理工大学大学生创新创业计划(编号:X2015052;X2016121))资助项目
Author NameAffiliationE-mail
Shen Yangli Medical Research Center,North China University of Science and Technology 1256461093@qq.com 
Sun Hao Medical Research Center,North China University of Science and Technology  
Cai Wenchen Medical Research Center,North China University of Science and Technology  
Zhang Pinzheng Medical Research Center,North China University of Science and Technology  
Wang Xuying Medical Research Center,North China University of Science and Technology  
Shang Yuhan Medical Research Center,North China University of Science and Technology  
Shi Lu Medical Research Center,North China University of Science and Technology  
Guo Zhiyi Medical Research Center,North China University of Science and Technology guozhiyi@ncst.edu.cn 
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Abstract:
      the detection of nuclear acid polymorphism has important role in basic research and clinical application. The popular methods are PCR based on probe. Probe with cross-reactivity limits the application of traditional real-time quantitative PCR to quantitate allelic transcripts. Digital (d)PCR may overcome cross-reactivity defects, but available dPCR mchines lack discrete optical channels which limits the detection of more than two molecules. Colon cancer associated transcript 2 (CCAT2) is a non-coding transcript, encompassing rs6983267 SNP site. Here, we report a method based on two channel droplet digital PCR to quantitate three CCAT2 polymorphism in one reaction. We designed one primer pair and three hydrolysis probes including a reference and two competing probes. We successfully discriminated between the three CCAT2 transcript clusters. We took advantage of the cross-reactivity to provide a white space for more visible distinct clusters, although innate rain was detected. Labelling the reference probe same with cross-reactivity probe resulted in more distinct clusters than if the reference probe was labelled in the same way as the probe without cross-reactivity.
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