DNA extraction protocol using Chelex-100 is cheap and very useful technique. However, the protocol varies between lab to lab, or people to people.
My lab is fungal taxonomy lab, so DNA is frequently from lab-grown fungal colony.
Currently in my lab, the protocol is:
1. Prepare 10% of Chelex-100 solution with ddH2O, stored at 10'C convective fridge.
2. aliquote 300λ of Chelex sol. to 1.5mL tube, harvest fungal colony (about (0.5mm)^3), from media, mix it by inverting or vortex mildly.
3. Pre-heat samples 56'C with Heating block for 15 min.
4. Load the tubes into floater, boil at 100'C for 10 min.
5. Vortex tube vigorously.
6. Again, Boil the tubes at 100'C for 8 min.
7. Centrifuge at 101g, 4'C, for 10 min.
So far there were no problems yet, but I'm looking for ways to improve yield or purity of the DNA.
Would this protocol be changed or fixed?
Waiting for the Answers from the best :)
you can test both of Chelex and Te buffer as:
mix chelex 100 150 ml and 50 ml TE buffer
add sample and squelch and crush it in mix of two liquids for 30 second
after put in PCR in 95 c for 5 min
after take 4 ml for PCR reaction
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