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。
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.
出处
《生物信息学》
2010年第2期139-141,共3页
Chinese Journal of Bioinformatics
基金
863计划项目(2006AA020903)
