Thread subject: Diptera.info :: DNA extraction from samples

Posted by RobertOBaggio on 09-04-2014 19:14
#1

Hello folks,
I am hoping to extract DNA from samples collected from Malaise traps. Whilst I have been warned about using 70% IMS in my collection bottles due to DNA denaturation, I was hoping that someone might be able to comment on one proposed way around of having to use ethanol in my collecting heads:

If the insects are trapped in IMS (70%) and transferred weekly to ethanol, then the DNA should not degrade very much. I have read that IMS degradation occurs over periods of weeks rather than days.

Cheers.

Posted by jonas on 10-04-2014 14:03
#2

Robert,
70% ethanol for DNA extractions, from malaisetraps works perfect. No worries.

In a barcoding project on Empidoidea we got a very high success rate for COI. All samples were malaisetrap samples collected directly EtOh, stored for two weeks at 20° (in the dark!), and then identified and each specimen put in a separate vial in 96% EtOh.

I have no experience with IMS - but why not collect directly in EtOh?

Please, for details check our publication: http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3890682/
Cheers

Posted by RobertOBaggio on 10-04-2014 14:06
#3

Thanks Jonas,

The issue with EtOh is cost. I will be running 24 Malaise traps for 5 months (weekly collections). IMS is around 5 times cheaper than EtOH. I will defo read your paper. Many thanks for the reply. I hope to do some barcoding myself so fingers crossed!



Posted by ChrisR on 12-04-2014 12:08
#4

Where will you be collecting? Remember to keep any tachinids :D

Posted by libor on 12-04-2014 17:21
#5

Roberto, I would highly appreciate Anisopodidae, Rhagionidae, and Athericidae. Is this material from your traps free?
Thanks
Libor