We used the DNA Clean & Concentrator-100 (D4030) from Zymo Research to clean and concentrate our 100 µg DNA. Unfortunately, our yield was only 8%.
We initially believed that, due to our DNA being over 10 kb, it was not eluted properly from the column. However, upon checking the DNA in the flow-through and wash samples at each step, we discovered that most of our DNA did not bind to the column. Specifically, 90% of our DNA existed in the flow-through. When we applied the flow-through containing this DNA to another company's DNA binding column, most of the DNA bound well to the column. Consequently, we suspect there is a problem with the DNA binding column of the Zymo kit.
Has anyone had a similar experience? If so, how did you resolve the problem?
How much DNA do you add per column? What was the sample/buffer you used? How do you measure DNA cc?
I have been using that kit for years and the yield is consistently 50-80%
I have a similar problem. Obtaining 4% of original midiprep plasmid dna from this kit. Any updates on fixing this issue?
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