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主题:【原创】这是真的吗?Maxipreps concentration=4163.1ng/ul?! -- 大鹏翔宇

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家园 Small questions/suggestions

Nice figure.

I have several questions/suggestions, though.

1. It would be more informative if you could label the DNA ladder, I tried to guees the size but I could not correlate the sizes.

2. You may want to try to load less sample. It looks to me that you've loaded more than you need - or may be it's just the result of photographying. As a matter of fact, you might be able to distinguish the two bands in lane 5/9 if you chose carefully the amount of sample loaded and the gel concentration.

3. Load the empty lane with loading buffer (prefereably the same ionic strenght as the samples) might help to straighten the running lanes.

4. Lane 9 shows you have not completely digested the sample. The tails in Lane 7-9 may indicate that you have some impurities in your phRL-SV40 vector preparation. This might explain partially why you got the high yield - you could get some E coli genome DNA with your plamid. However, this is just my guessing.

There is a step after you broke the E coli where you need mix that gelly stuff with buffer. You need to mix them gentlely, otherwise, you might break some genome DNA, which will not be separated with centrifugation since it will not precipitate. In another word, it will go with your plasmid sample.

[The above are just my own opinion. We can discuss them if you have different thoughts.]

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