I will be extracting trypanosome DNA from mid-guts of tsetse flies for LAMP analysis. Kindly advice on protocol to adopt for maximum yield and benefits of Chelex method over the regular method. Thanks in advance!
Hi Rolayo,
In my experience, Chelex based methods are great for extracting DNA from small tissue samples or swabs (basically when you have very little starting material). It is relatively cheap when compared to extraction kits. It isn't the cleanest extraction method, so additional cleanup step may be required, but I have seen good results with Chelex extractions in most down stream applications (PCR, qPCR, and NGS). I have been following Caquet 2012 and adjusting volumes based on tissue size and desired final elution volume.
Quick tips:
- Chelex beads are inhibitors. Do not transfer any Chelex beads into your LAMP assay.
- Because 10% of the reaction volume is Chelex, you will have a lower elution volume than expected. Bump up your extraction volume to account for the Chelex beads.
Article Chelex without boiling, a rapid and easy technique to obtain...
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