| 基于RefSeq的人类基因荧光定量PCR引物库的构建 |
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| 引用本文： | 周贵良,洪来法,梁敏玲,郑柳城,陈转贤. 基于RefSeq的人类基因荧光定量PCR引物库的构建[J]. 生物信息学, 2010, 8(2): 139-141 |
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| 作者姓名： | 周贵良 洪来法 梁敏玲 郑柳城 陈转贤 |
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| 作者单位： | 广州复能基因有限公司,广州,510663 |
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| 摘 要： | 利用NCBI提供的RefSeq序列,通过BLAT、Sim4和自主开发的剪接比对程序Ealter1.0对人类RefSeq转录本进行外显子预测,根据预测的外显子信息,采用自主开发SYBR Green I Real Time PCR引物设计程序E-qPCR-Design1.0高通量设计21,118对SYBR Green I Real Time PCR引物,同时选取5000条基因进行SYBR Green I Real Time PCR引物验证,95.92%的基因引物取得良好效果,1.64%的基因引物产生引物二聚体,1.08%的基因引物有非特异性扩增,通过生物信息技术分析与实验验证,建立了基于RefSeq的人类基因荧光定量PCR引物库。
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| 关 键 词： | RefSeq转录本数据库 实时定量PCR |
Construction of human gene SYBR Green qPCR primer database base on RefSeq sequence |
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| ZHOU Gui-liang,HONG Lai-fa,LIANG Min-ling,ZHENG Liu-cheng,CHEN Zhuan-xian. Construction of human gene SYBR Green qPCR primer database base on RefSeq sequence[J]. Chinese Journal of Bioinformatics, 2010, 8(2): 139-141 |
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| Authors: | ZHOU Gui-liang HONG Lai-fa LIANG Min-ling ZHENG Liu-cheng CHEN Zhuan-xian |
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| Affiliation: | ( Guangzhou FulenGen, Co., LTD ,guangzhou 510663,China) |
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| Abstract: | According to NCBI RefSeq database, a large scale exon prediction analysis for maping all of RefSeq records of human to hu- man genome sequence database was further performed using BLAT, Sim4 and a homemade program, Ealterl. 0. According to the exon prediction information, we designed 21,118 pairs SYBR Green qPCR primer by E -qPCR -Designl. 0, and 5 000 genes selected SYBR Green I Real Time PCR primer validation, 95.92 % of the primers to obtain good results, 1.64% of the gene primers generated primer dimers, 1.08% of genes have non - specific primer amplification, through bioinformatics analysis and experimental verification, based on Green I Real Time PCR primer database. |
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