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wansanlian

金虫 (小有名气)

[求助] CTAB提取液配制时加入了PVPP怎么不溶解啊? 已有1人参与

我在配制CTAB提取液的时候加入了PVPP,可是一直都不溶解,混了一天一夜了,还是乳白色的,拿去高温灭菌也还是一样的,都不溶解,有没有人知道是怎么一回事啊?
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junminh

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【答案】应助回帖

感谢参与,应助指数 +1
分开溶,再合并到一起。乳白色说明没有完全溶解,CTAB没有完全溶也有可能。放在80-100度,2H,应该OK
2楼2012-09-23 08:38:45
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xiaoyi855

新虫 (初入文坛)

引用回帖:
2楼: Originally posted by junminh at 2012-09-23 08:38:45
分开溶,再合并到一起。乳白色说明没有完全溶解,CTAB没有完全溶也有可能。放在80-100度,2H,应该OK

我也遇到类似的问题了,分开溶还是溶解不了。加热水、高温80度,灭菌等等都试过了,就是不溶解。
3楼2018-10-19 10:04:13
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xiaoyi855

新虫 (初入文坛)

【答案】应助回帖

引用回帖:
2楼: Originally posted by junminh at 2012-09-23 08:38:45
分开溶,再合并到一起。乳白色说明没有完全溶解,CTAB没有完全溶也有可能。放在80-100度,2H,应该OK

When guanidine based method repeatedly gave negative results, we switched over to CTAB based method, and appropriately modified the protocol as per the requirements of RNA isolation from a phenolic rich plant tissue. The improved CTAB protocol described here efficiently eliminated most of the interfering molecules and yielded translucent and water soluble RNA pellets. CTAB buffer, as described by Chang et al. [9] was used for this purpose. CTAB was used as a cell disrupting agent, supplemented by insoluble PVPP instead of soluble PVP as it is incompatible with phenol extraction and would have obstructed RNA precipitation [10]. PVPP as an alternative is insoluble, cross-linked form of PVP with high molecular weight which is chemically inert and forms complex with polyphenols through hydrogen bonds, allowing them to beseparated from nucleic acids [11].

是不是可以理解成配制完含有pvpp的CTAB就是浑浊状态的?可以直接用于提取RNA?

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4楼2018-10-19 15:32:12
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