I attached the gel we got after doing plant DNA extraction. We follow the next steps and now we're supposed to figure out what went wrong and what can we do to optimize our method. Do you see something we did wrong?
Cut a sample disc using the Eppendorf tube.
Macerate the sample at room temperature (without buffer) for 15 seconds.
Add 400 uL of extraction buffer and agitate with vortex for 5 seconds.
Centrifuge at 13000 rpm for 1 minute.
Take 300 uL of the supernatant and transfer it to a new Eppendorf Tube.
Add 300 uL of isopropanol and let it rest for 2 minutes.
Centrifuge at 13000 rpm for 1 minute.
Redissolve the pellet in 100uL of buffer TE 1X.
Ahhh, then you didn't extract DNA at all... Try the CTAB-method or a kit!
Isopropanol pellet. It's very easy to displace. Most likely you lost it when removed the supernatant. I would recommend using a kit with a column purification to avoid the precipitation step. Alternatively, your homogenization procedure was not efficient enough and you started with very little DNA.
The attached image reveal, you has not DNA extract from your samples so I agreed with Nadine A. Gund, thus try to use a kit, before PCR, measured the obtained DNA by any tool available there in your lab
By the look for the ladder shape , it seemed there were a problems with the electricity power supply
Maybe no problem with the electricity power supply but I think the running time was too short! Cheers, Nadine
You didn't extract DNA. Try the CTAB-method.
Or modify your present method by avoiding vortex (you just invert tubes manually) & use double volune of isopropyl alcohol.
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