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Diptera.info :: General Diptera forums :: Methodology
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DNA extraction from samples
RobertOBaggio
#1 Print Post
Posted on 09-04-2014 19:14
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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.
 
jonas
#2 Print Post
Posted on 10-04-2014 14:03
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Joined: 06.05.07

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
 
RobertOBaggio
#3 Print Post
Posted on 10-04-2014 14:06
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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!


 
ChrisR
#4 Print Post
Posted on 12-04-2014 12:08
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Location: Reading, England
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Where will you be collecting? Remember to keep any tachinids Grin
Manager of the UK Species Inventory in the Angela Marmont Centre for UK Biodiversity at the Natural History Museum, London.
 
http://tachinidae.org.uk
libor
#5 Print Post
Posted on 12-04-2014 17:21
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Location: western Bohemia
Posts: 1124
Joined: 30.05.09

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