Diptera.info :: General Diptera forums :: Methodology
Who is here? 1 guest(s)
 DNA extraction from samples
|
|
| RobertOBaggio |
Posted on 09-04-2014 18:14
|
|
Member Location: Posts: 43 Joined: 31.01.14 |
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 |
Posted on 10-04-2014 13:03
|
|
Member Location: Posts: 351 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 |
Posted on 10-04-2014 13:06
|
|
Member Location: Posts: 43 Joined: 31.01.14 |
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 |
Posted on 12-04-2014 11:08
|
 Super Administrator Location: Reading, England Posts: 7706 Joined: 12.07.04 |
Where will you be collecting? Remember to keep any tachinids 
Manager of the UK Species Inventory in the Angela Marmont Centre for UK Biodiversity at the Natural History Museum, London. |
 |
|
| libor |
Posted on 12-04-2014 16:21
|
|
Member Location: western Bohemia Posts: 1294 Joined: 30.05.09 |
Roberto, I would highly appreciate Anisopodidae, Rhagionidae, and Athericidae. Is this material from your traps free? Thanks Libor |
|
|
|
| Jump to Forum: |