efficiency of Qiagen Kit

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huyenkobe
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efficiency of Qiagen Kit

Post by huyenkobe » Fri Jan 07, 2011 4:19 am

Hi everyone,
Sure that many of you use the kit of Qiagen. I am using the MiniElut purification Kit which mention that the efficiency of recovery DNA from gel can reach up to 80% by using this Kit. Actually, I repeat so many times and the best I had only around 30%. Anyone know how to increase it. I tried many ways, add sodium acetate, increase the time to solubilize gel ... but nothing improves. It causes the lost of product, when my boss does not want to increase the amount of product but only try how to improve the efficiency. I have to begin with 5microgram of plasmid, and after purification, usually I have nothing left. Anyone know how to solve the problem.

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canalon
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Post by canalon » Fri Jan 07, 2011 5:22 am

What do you elute with? Do not use water but the elution buffer (or 10mM Tris pH8).
I warm the column for 10 minutes at 50ºC opened in a dry block to improve removal of EtOH and add warmed buffer (50ºC too) then keep 5 minutes in the dry block before spinning. I know that in some protocols based on the same technique, you pipette the elution and reload it on the column to improve slightly the recovery.

And by the way, there are many other kit as good, if not better for some applications, and wayyy cheaper than QIagen. Some are crap though.

Good Luck
Patrick

Science has proof without any certainty. Creationists have certainty without
any proof. (Ashley Montague)

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