三七是我国重要的药用植物之一,又名田七,人参三七等,为五加科人参属植物,是我国传统名贵药物,自古就有“止血神药”的美称。距今已有400多年的应用历史,在国内外久负盛名。三七主产于云南文山。品种缺乏是三七种植存在的重要技术问题,育种标记和方法则是困扰三七育种的重要问题,这与我们对三七的遗传基础缺乏了解有关。
研究发现,三七是在9100万年前与茄科植物马铃薯和辣椒分化开来,三七与四倍体的人参属植物人参和西洋参有9383个共同的基因,976个特有的转录本.,基因组测序的完成将为三七全基因组选择育种、功能基因的发现等提供了基础,也有利于三七产业关键技术问题的解决。
三七样品中提取DNA,中国农业大学农学院之前采用研钵手工研磨的方法,在研钵内倒入液氮,在冻干状态下快速研磨,研磨过程中样品不易研磨,整体研磨样品不充分,影响实验效果,因此选择上海净信冷冻组织研磨仪。
实验材料和器具:
样品:三七
工具:净信冷冻组织研磨仪(JXFSTPRP-CL-24)
耗材:钢罐,研磨珠、液氮、剪刀、镊子等
实验步骤:
1、准备钢罐2个,研磨珠若干颗;
2、取三七组织剪成小块与研磨珠一同放入钢罐中,将盖子拧紧;
3、将钢罐置于装有液氮的容器内,液氮容量没过钢罐即可,冷冻若干分钟,取出时戴手套或使用镊子夹取避免液氮冻伤;
4、将适合器放入研磨仪器中固定好,然后开机并将制冷功能打开进行预冷
5、将浸泡过液氮的钢罐放置适合器内,样品尽量对称放置,以保证研磨时的受力平衡,旋紧垂直定位螺旋,将安全锁扣紧。设置参数,程序结束后即可得到破碎粉末;
6、研磨完毕后请关闭电源,将安全锁归位;
7、完成后续 DNA 提取步骤。
注意事项:
1、每支钢罐内的样品量一般不超过罐体体积的三分之二,样品量过多会影响研磨效果。
2、样品尽量对称放置,以保证研磨时的受力平衡。
3、样品夹取过程中戴手套及使用镊子夹取避免液氮冻伤。
实验过程、结果图示:
实用仪器介绍:
此款仪器为上海净信实业发展有限公司生产的全新全自动样品冷冻研磨仪,它是一种特殊的、快速的、高效率的、多试管的一致系统。可以在较短的时间内、 低温状态下完成对各种样品的研磨、粉碎、混合及细胞破壁,以保留蛋白质活性。
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