缓冲液离子浓度对新型PNA-LSAW基因传感器杂交效应的影响

Effects of ion concentrations of buffer on hybridization efficiency with new style PNA-LSAW gene sensor

目的探讨PBS缓冲液的离子浓度对构建的肽核酸(PNA)漏声表面波(LSAW)基因传感器杂交效应的影响。方法将1.0μmol/L的PNA探针固定在LSAW基因传感器表面,分别加入10、20、50、100mmol/L的PBS缓冲液,观察不同离子浓度的PBS缓冲液对PNA-LSAW基因传感器杂交效应的影响,记录分析传感器相位的变化值及反应所需的时间。结果当钠离子浓度从10mmol/L增加至100mmol/L时,传感器的相位变化呈先升高后降低的趋势,以20mmol/L为分水岭。而杂交平衡时间增加显著。结论离子浓度极大地影响了杂交反应的时间,低盐离子浓度更有利于提高PNA和靶序列的杂交速率,高盐离子浓度则会大大降低PNA的结合速率。离子浓度为20mmol/L时杂交最为适宜。

Objective To investigate the effects of ion concentrations of PBS buffer on hybridization efficiency with peptide nucleic acid（PNA） leaky surface acoustic wave（LSAW） gene sensor. Methods 1.0umol/L PNA probe was firstly immobilized on the surface of LSAW gene sensor. Then it was hybridized with corresponding target sequence when ion concentrations of PBS varied from 10mmol/L to 100mmol/L,and the changes of phase and equilibrium time of reaction were recorded and analyzed. Results Phase changes of LSAW sensor were increased firstly and decreased laterly when ion concentrations varied from 10mmol/L to 100mmol/L,and 20mmol/L was the watershed. However,equilibrium time was increased significantly. Conclusion Hybridization time was effected greatly by ion concentrations. Hybridization rate of PNA reacting with target sequence was improved by low ion concentration,while it was reduced significantly by high ion concentration. 20mmol/L of the ion concentration was optimal.