實(shí)驗(yàn)?zāi)康?/strong>
研磨破碎腫瘤組織細(xì)胞(如胰腺、肺、肝等器官的腫瘤組織),后續(xù)提取其RNA。
實(shí)驗(yàn)原理
通過研磨珠撞擊腫瘤組織細(xì)胞,使其細(xì)胞膜破裂,其內(nèi)的RNA暴露。
實(shí)驗(yàn)材料和器具
樣品:腫瘤組織細(xì)胞
儀器:高通量組織研磨儀(上海凈信,Tissuelyser-48)
耗材:離心管,研磨珠(上海凈信)
實(shí)驗(yàn)步驟
1.取組織塊解凍,用生理鹽水洗去血污,剪取少許組織,剪小放入離心管中;
2.加入研磨珠,加入TES混勻,再加入SDS,蛋白酶K,充分混勻;
3.離心管放入研磨機(jī)中,按要求設(shè)置好參數(shù),程序結(jié)束即可得到破碎原液;
4.完成后續(xù)RNA提取步驟。
實(shí)驗(yàn)結(jié)果
圖1 胰腺腫瘤組織RNA提取
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