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研究适用于昆仑雪菊的ISSR-PCR反应体系,建立ISSR分子标记技术评价昆仑雪菊生态多样性体系。采用改良CTAB法、CTAB区室法、改良CTAB区室法、试剂盒法提取昆仑雪菊的新鲜叶片、干花以及种子的DNA;以827为引物,研究浓度、循环数、退火温度对ISSR-PCR反应体系的影响,以不同实验材料检测其适宜性。结果表明,昆仑雪菊新鲜叶片、干花以及种子的DNA的最佳提取方法为改良CTAB区室法,ISSR-PCR反应体系的最适引物浓度为1.0μmol·L-1,退火温度为54.6℃,循环数为30。ISSR-PCR反应体系适用于评价昆仑雪菊的生物多样性。
The ISSR-PCR reaction system suitable for Kunlun Xueju was studied, and the ISSR molecular marker technique was used to evaluate the Kunlun snowdrop ecological diversity system. Using the modified CTAB method, the CTAB compartment method, the modified CTAB compartment method and the kit method, the DNA of fresh leaves, dried flowers and seeds of Kunluosia chrysanthemi was extracted. Using 827 as the primer, the concentration, number of cycles and annealing temperature were studied. Reaction system to test the suitability of different experimental materials. The results showed that the optimal extraction method of DNA for the fresh leaves, dried flowers and seeds of Kunlun snow chrysanthemum was improved CTAB compartment method. The optimal primer concentration of ISSR-PCR reaction system was 1.0 μmol·L-1, the annealing temperature was 54.6 ℃, The number of cycles is 30. The ISSR-PCR reaction system is suitable for evaluating the biological diversity of Kunlun Xueju.