有去除细胞碎片的好方法吗？因为是原代分离的细胞，所以碎片比较多，想送去RNA-Seq

微笑流滢

因为是原代分离的细胞，所以碎片比较多，想送去RNA-Seq，细胞碎片太多会不会影响RNA质量？有没有去除细胞碎片的好方法？

lchanlon

The cell debris are in the RNA samples? For RNA-seq, before library preparing, RNA need to be qualified first. Typically, using Agilent Bioanalyzer to do quality control. The RIN (RNA Integrity Number) should be greater than 8. Based on the kit you are using, at least 100ng of total RNA is needed. If your RNA RIN is good enough and you have much more amount of RNA than 100ng. Cell debris is not a problem.

lchanlon

The cell debris are in the RNA samples? For RNA-seq, before library preparing, RNA need to be qualified first. Typically, using Agilent Bioanalyzer to do quality control. The RIN (RNA Integrity Number) should be greater than 8. Based on the kit you are using, at least 100ng of total RNA is needed. If your RNA RIN is good enough and you have much more amount of RNA than 100ng. Cell debris is not a problem.

misser217

splinkerette

可以传代的话直接消化传代，半小时后差不多细胞能贴壁，用显微镜观察确认后，更换培养基，基本应该能去除细胞碎片

bubble_w

回复 微笑流滢 的帖子5 ?. W4 i2 k4 b1 r$ T0 G

- p' @, V  n1 j原代分离的细胞有很多细胞碎片和组织碎片，或者一些组织中胶原含量高，也容易去除不净。如果够条件就做FASC纯化一下，如果不行，就贴壁培养。贴壁培养的好处是细胞纯度高，状态好，但是仅限个别细胞种类。还可以用差速沉降的方法，加入大量的新鲜培养液，重悬细胞沉淀，静置3-5min，去除上清，多次重复，也可以部分减少原代细胞中的细胞碎片。

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: m, e2 @. g' qThank you ,your english is so good . I want to make friends with you

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