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nalanyuhang

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[求助] 水氯仿甲醇提取组织代谢物 已有1人参与

实验中要用CE/MS测肿瘤组织中代谢物,代谢物提取过程如下:组织加20ul/mg甲醇匀浆后,加水,氯仿(体积比5:2:5),离心取上清。。。。。。现在想知道的是加水、甲醇、氯仿的目的各是什么?离心后上清的水相中有甲醇吗,还是甲醇在氯仿相中?文献中只要水溶液是因为小代谢物大多水溶吗,这样只要水相结果会精确吗?                在线等大家一起交流指导,感激不尽!!
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xianl13

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2楼2017-10-29 12:11:53
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僵屎

新虫 (初入文坛)

引用回帖:
2楼: Originally posted by xianl13 at 2017-10-29 12:11:53
请问你加入提取液后离心后,组织是在底部还是在中间呀?我今天在组织中加了提取液(甲醇:水:氯仿=5:2:2)然后离心,组织悬在中间,底层应该是氯仿而且颜色是透明的,上层上清是黄色的。我记得之前做过组织是沉淀在 ...

Samples were vortexed for 1 min, left for 10 min on ice
for partition and centrifuged (2,0009g, 10 min, 4 C) to
obtain a clear biphasic mixture, upper aqueous/polar phase
and lower organic/apolar phase. The upper polar phase
was collected and the lower organic layer was carefully
recovered using a positive-displacement Microman pipette
(Gilson, Middleton, WI, USA), without disturbing the
layer between the two phases containing protein and small
tissue debris.

组织和蛋白沉淀在中间层
好人有好报
3楼2018-09-25 16:04:07
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2551359343

新虫 (初入文坛)

你好,请问你问题解决了吗,我也想问这个问题,可以向你请教吗
4楼2018-12-21 20:55:20
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