摘要
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.
出处
《生物技术》
CAS
CSCD
2007年第3期42-44,共3页
Biotechnology
基金
沈阳市科学事业费专项资金资助项目("玉簪花组培快繁技术研究"
编号:2005020117)