I use DNA barcoding to look for “fish fraud” in my undergraduate biology lab class. It’s a HUGE course (~2000 students per year). The DNA extraction protocol I’ve been using works well except it takes way too much time (about 2 hours for the students). Anyone know a faster protocol to get DNA out of raw fish? And we don’t have a fume hood so can’t do phenol chloroform preps.
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Hi Katie
Will you be using PCR downstream to confirm ? If yes there are multiple ways to get this done and that is not dependent on the total DNA integrity but PCR based readiness of your sample. A simple protocol in that case would be based on:
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Collecting a Small tissue sample from Fish
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Homogenize and add a Lysis buffer ( usually Tris ph 8.0, SDS & EDTA supplemented with Proteinase K).
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Incubate for 15 min
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Inactivate Proteinase K by heating at 95 C
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Centrifuge to remove any debris and then collect 1-5 microlitre sample and Proceed with PCR.
This should be sufficient for your work/ Demonstration
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Thank you!!
Yes, it’s for DNA barcoding. I’ll have to test it out in Summer. Having the students do DNA extraction and PCR in one lab will really speed up their project
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Hi Katie
In this case it should work well for you. The protocol is rapid, cost effective and doesnt require specialized/ expert handling. I suggest running a few Pilot experiment samples to test before starting with a large batch
Let us know If you’d require help at any step ahead.
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Hi Katie
Were you able to do a Pilot run? How were the results? In case you face any hurdles you can share the images of your gel run and we can figure it out further.
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I’ll have time to test it out over summer, I’ll let you know what I find out!
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