5种毁灭你琼脂电泳的方式
发表于274 天前 实用 评论数 4 ⁄ 被围观 1,350+

找生物-分享有趣、实用的生物资源!今天送出的是一个5种毁灭实验方式系列的一个,主角是琼脂糖电泳。原文比较简单,找生物对大条目进行了简单翻译,具体文章自己看吧,文中列举的这五种方式,各位童鞋您遇到过吗?找生物是经历过!呵呵,各位童鞋也分享一下你的遭遇吧!

琼脂糖凝胶电泳

1. 使用水配胶或把水当做运行缓冲液

Agarose gels are cast and run using TAE or TBE buffer. Since both of these buffers are clear, it’s easy to mistake them for water. If you do use water though, your gel will melt shortly after applying charge to the gel box – say goodbye to your samples!

2. 忘记加EB

In a rush, it’s easy to forget adding ethidium bromide to your agarose before casting the gel. Without it, of course, it will be impossible to visualize your DNA. You can rescue an unstained gel by shaking it in an ethidium bromide solution after it’s done running; but then you have a large volume of waste to dispose of.

3.使用错误的琼脂浓度(或琼脂类型)

The standard percentage of agarose used to run a DNA gel is usually around 1.0%. Using a higher agarose percentage enhances resolution of smaller bands; conversely, using a lower agarose percentage allows the smaller bands to run through the gel quickly, giving you better resolution and separation of higher molecular weight bands. If you use the wrong percentage, it can be difficult to visualize your bands reliably. Be careful when you start decreasing the percentage of agarose in your gel, as they become weak and much more prone to breakage.

Low melting point agarose is used for specialized applications, such as in-gel ligation. These gels are very “soupy” and fragile, even at comparable percentages, so make sure you grab the right reagent before mixing up a batch of agarose.

4. 接反正负极

One of the most frustrating mistakes you can make is to accidentally switch the leads on your power source, so that the gel runs backwards. Since the wells are so close to the end of the gel, you will most likely lose all of your samples before you catch the mistake. If you see your bands heading in the wrong direction, though, just switch the leads and run it back in the other direction.

5. 在去照相的途中把胶给弄丢

Seriously – don’t let this happen to you.这个是最严重的事情了,不要让这种事情发生你身上,找生物悲怆了!

源文链接 <http://bitesizebio.com/articles/5-ways-to-destroy-your-agarose-gel/>

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