I want to examine insulin like peptides in crickets, however no primers exist for ILP's in crickets nor any methods of detection. They exist in the drosophila literature- I just am at a standstill on trying to figure out how I could do this without spending a fortune.
hi
It may help you !
http://www.ent.uga.edu/pubs/brown_riehle_molecular.pdf
There is a cricket transcriptome around. Did you check if the precursors are present there?
http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0061479
Ah, if only I was using Gryllus- I'm using Acheta domesticus as my model organism. Thank you though!
What kind of techniques were you considering in the first place? Is mass spectrometry an option?
Otherwise, try to collect some orthoptera sequences, find some conserved spots and design degenerate primers?
I don't believe it is and time is a factor in this as well. Since I'm relatively new to the world of neurobiology in arthropods- is it possible that since both A.domesticus and G.bimaculatus are in the same sub-family of Gryllinae, primers designed for G.bimaculatus might work in A.Domesticus?
Thank you so much for your help and brainstorming!
You can always give it a try, with not-too-stringent PCR settings. You may get many bands, but you'll also have an idea about the expected fragment size, so that could work...
I'd definitely check more than one orthoptera sequence to see where you have the highest likelhood of conserved stretches. Once you have a short fragment, you can use RACE.
Just out of curiosity: how are you getting on with your cricket peptides?
Happy new year :)
Rik
Hi Rik!
Happy New Year to you as well =). We've now put that aside to answer more immediate questions with my time left (1 year) at my institution.
- Badges
- Science topic
- Similar topics
- Biological Science
- Neuroscience