鸭茅ISSR反应体系的建立及引物筛选

Establishment of ISSR Reaction System of Primer Filtration Dactylis Glomerata

以鸭茅基因组DNA为模板，对ISSR反应体系中的一些重要参数进行摸索和优化实验，建立了一套鸭茅ISSR分子标记的最优化反应体系，同时筛选出了12个适于鸭茅基因组DNA且PCR扩增效果较好的ISSR引物。鸭茅ISSR分子标记的20μL最优化反应体系中，最终浓度：Mg^2＋为1．6mmol／L、dNTP为250mol／L、Taq酶为1．0 U、引物浓度为0．25μmol／L、模板DNA量为100ng。

By using the genomic DNA from orchardgrass （Dactylis glornerata L.）, some essential factors that might affect the result of ISSR assay were tested and compared. Then an optimal ISSR reaction system for Dactylis glornerata was established. That is, in a volume of 20 μ L amplification reactions system, there are 1.6 mmol/L Mg^2＋, 250 μ mol/L DNTP, 1.0 U Taq DNA polymerase, 0.25 μ mol/L primer, 100 ng template DNA. After the establishment of ISSR Reaction System, twelve good primers for Dactylis glornerata were obtained after priner screeing.