关于ISSR反应弥散背景消除方法的几点讨论

Discussion on Eliminating the Smear of ISSR Reaction

目的：讨论消除ISSR反应弥散背景的一些方法。方法：对于不同的反应条件,分别进行梯度试验,包括：退火温度（45℃、47℃、49℃、50℃）、不同引物浓度（0.2μmol/L、0.4μmol/L、0.6μmol/L、0.8μmol/L）、不同模板浓度（50ng、100ng、200ng）及不同纯度。结果：梯度试验结果如图1~10所示。退火温度47℃,引物浓度0.6μmol/L,模板浓度100ng左右,纯度高的模板,在这些条件下得到的电泳图谱比较清楚。结论：导致弥散背景产生的主要原因有：过低的退火温度;不适当引物浓度;较高的模板浓度;还有模板纯度不高。

Objective：The paper mainly discussed on the method of eliminating the smear of ISSR reaction. Methods：The ladder experiments were designed that included the different anneal temperature （45℃ ,47℃,49℃ ,50℃ ）, prime concentration （0.2μmol/L,0.4μmol/L,0.6μmol/L,0. 8μmol/L）, sample concentration （50ng,100ng,200ng） and sample purity. Results：The results of the ladder experiments were illustration on the pictures from 1 to 10. Conclusion：The smear was induced by too low anneal temperature, not suitable prime concentration,high template concentration and not dean template.