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alice keer新蟲(chóng) (初入文壇)
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[求助]
真菌菌絲提取及電泳圖求解釋?zhuān)? 已有1人參與
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用CTAB提取法提取真菌DNA,但是抽提DNA過(guò)程中只用了氯仿/異戊醇(24:1),沒(méi)有使用Tris飽和酚抽提。在1%瓊脂糖凝膠電泳中檢測(cè)有無(wú)條帶,如下圖所示,請(qǐng)教各位大神:這是什么原因? 急需,感激不盡。。。。。。 20151216DNA(SHEN).jpg |
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The 4th step would be the problem. The order is different from my CTAB protocol, which is "Add 0.5 volume 5M NaCl and mix properly. Then add 2/3 volume ice-cold isopropanol and leave at least 20minutes (up to overnight) at -20 degrees Celsiu." It must be pointed out that once DNA and polysaccharides co-precipitate together, it becomes much more difficult to separate them than prior to precipitation. By adding 0.5 volum 5M NaCl to the CI-extracted aqueous solution just prior to precipitation by isopropanol, it is effectivly separate the DNA from polysaccarides. In my opinion, this step is the key to CTAB protocol. |
新蟲(chóng) (初入文壇)
鐵蟲(chóng) (小有名氣)
| 建議你把你做的全部步驟寫(xiě)上來(lái),理論上我們CTAB提都是百發(fā)百中,沒(méi)有提不出來(lái)的~~~而且沒(méi)有酚應(yīng)該也不至于啥都沒(méi)有,另外跑基因組DNA你要用HINDiii的maker或者其他大于10k的maker,我猜,你第二泳道上面那個(gè)弱弱的有可能是,但是5000的maker看不出來(lái)那個(gè)帶多大 |
新蟲(chóng) (初入文壇)
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