对于一些特别坚硬或韧性比较强的样品,如:皮肤、硬骨骼、软骨、血管、结缔组织、种子等,使用普通机械研磨方法很难即保证研磨破碎效果又保证样品一直处于低温状态。传统的手工研磨方法虽然能二者兼顾,但是当样品数量比较多时,手动研磨非常的费时费力,而且经常存在浪费样本、样本间交叉污染的可能,影响后续实验。如果提取RNA、酶等对温度比较敏感的物质,使用液氮进行预处理,保证样品在破碎过程中一直处于低温是非常重要的,上海净信已经成功解决这一问题。
使用上海净信Tissuelyser全自动组织快速研磨仪进行组织细胞的破碎时,将样品用液氮进行预冷冻,这样既能保证样品的破碎效率又能保证样品一直处于低温状态。以血管和植物叶片为例,具体操作方法如下:
血管:
取新鲜血管,用剪刀剪成若干小块,然后放入离心管中,再加入净信研磨珠,盖上管盖。将整个试管放入液氮中进行冷冻几分钟或直到液氮不沸腾,然后放入机器中处理几十秒钟。取出样品管可见样品被破碎成乳白色粉末,然后加入提取缓冲液,可以进行接下来的后续提取,加入缓冲液之前能保证样品温度一直为-20℃以下。
叶片:
取植物叶片,清洁叶片表面后放入离心管中,加入几颗净信研磨珠,盖上管盖。将整个试管和适配器放入到液氮中冷冻几分钟或直到液氮不沸腾,然后取出放入研磨机器中机器处理几十秒钟。取出样品管可见叶片被破碎成绿色粉末,然后加入提取缓冲液,进行接下来的后续提取即可,加入缓冲液之前能保证样品温度一直为-20℃以下。
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