New England Biolab
Monarch DNA Gel Extraction Kit
T1020S
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The format is simple and appealing. The procedure fast. Unfortunately, in my hands, the DNA obtained is not clean. It always contamination at 220-230 nm, even higher than the pick at 260 nm. Probably Guanidine from the gel binding buffer, Therefore a DNA clean up step is necessary before using the DNA.
Gel purification of DNA for cloning
PCR products or plasmid DNA inside an agarose gel
The gel is cut and dissolved at 50C in a binding buffer containing Guanidine thiocyanate and sodium iodide. The mix is loaded into a column and then washed with an Ethanol-based buffer. Finally, the DNA is eluted in a very small volume (as low as 6 ul)
The DNA has to be cleaned again using the Monarch PCR & DNA Cleanup Kit
I have tried to optimize the extraction following all suggestions in this list from the manufacturer:
1 Melt your agarose completely
2 Minimize exposure to UV light
3 Use the smallest agarose plug possible
4 Ensure there is no ethanol in your eluate (adding an extra spin to dry)
5 Use the recommended amount of Dissolving Buffer
I also tested:
6 Adding an extra wash step
7 Washing with double the amount of wash buffer
8 Rinsing the sides of the column when adding the wash buffer. The contamination at around 220 is still there.
N/A
If you are stuck with this kit, you can add the extra step with Monarch PCR & DNA Cleanup Kit. You just elute from the Monarch DNA Gel Extraction Kit in 20 ul and proceed with the next protocol. Elute in only 6 ul. In the picture I attached, I combined 2 gel extractions of the same insert in this last step (note that the final concentration looks the same). I lost aprox. 50% of the DNA, but still had enough for the ligation.
The low elution volume
DNA obtained is contaminated
I wouldn't recommend this kit, until NEB makes a 2.0 version w/o the contamination problem. I a waiting for it! because the low elution volume is great!
Product Name: Monarch® DNA Gel Extraction KitSupplier Page