Methylation specific PCR of four genes Design of primers.doc

 PAGE \* MERGEFORMAT 11 Methylation specific PCR of four genes Design of primers Of: Lin Jiang, Yao Dongming, Qian Jun, Xu Wenrong, money shock, Zhu Zhaohui, Chen Qin, Wang Yali, Xiao Gaofei [Abstract] Objective: Methylation specific PCR (methylation specific PCR, MSP) primer design were discussed. Methods: Methyl Primer Express Software ?q Version 1.0 software, “methprimer” and “MSPprimer” on-line primer design online software and the light of foreign literature MSP primers CCAAT / enhancer binding protein (CEBP, CCAAT / enhancer binding protein (CEBP, fragile histidine triad (FHIT and death associated protein kinase (DAPK gene promoter for MSP expansion increase, then its MSP products were sequenced to compare the reliability of MSP primer design. Results: For CEBP, CEBP, FHIT, and DAPK gene promoter designed by MSP after MSP primers were needed to obtain the target band, However, after sequencing Methyl Primer Express and methprimer design CEBP, CEBP 1 of the MSP products amplified products as false positive, and MSPprimer designed CEBP 2 of the MSP product is the correct sequence. Conclusion: MSP primer design is to ensure the quality of MSP amplification critical success factor. [Keywords:] methylation specific PCR, primer design [Abstract] Objective: To study the primer design of methylation specific PCR (MSP). Methods: The Primer of MSP was designed using the software of Methyl Primer Express ?q Software Version 1.0, the online software of “methprimer” and “MSPprimer” to amplify CEBP, CEBP, FHIT and DAPK gene promoters.The MSP assay was established to detect the methylation status of these promoters.Then the MSP products were analyzed in electrophoresis and sequenced. Results: The expected bands were all obtained for all five primer sets. But sequencing revealed that amplification products using CEBP 1 and CEBP primers designed with Methyl Primer Express ?q Software and methprimer were false positive, amplification products using CEBP 2 pri