Feasibility of Guanidine Hydrochloride Method for ssDNA Library Construction of Osteosarcoma Aptamer
Objective Exploring feasibility of Streptavidin-Biotin(STV-Biotin)system coupled with Guanidine hydrochloride(G-HCl)method for construction of an osteosarcoma cell aptamer ssDNA library.Methods The candidate oligonucleotide library ssDNA was selectively bound to target cell(osteosarcoma cell U2-OS),and the oligonucleotides were amplified into double-stranded DNA(dsDNA)by PCR,dsDNA were labeled with biotin and combined with Streptavidin Sepharose(S.S)affinity chromatography.The target single-stranded DNA(ssDNA)library was prepared by G-HCl method and purified.The classical NaOH method was used for control.The single strand libraries prepared by the two methods were compared with osteosarcoma cells for compatibility and recovery.Results Screening library ssDNA obtained by G-HCl method have higher affinity to U2-OS and higher recovery of ssDNA than classical NaOH method.Conclusion STV-Biotin system coupled with G-HCl is a successful attempt for the preparation of osteosarcoma cell aptamer,which provides a new idea and method for the const-ruction of tumor aptamer ssDNA library.
国家科技期刊平台
NSTL
万方数据
