I'm trying to extract Bd fungal DNA from a cotton swabs after swabbing on frogs. I'm trying to get total DNA, but I'm not getting it. Right now I'm using lysis buffer containing EDTA, Tris HCl, Nonidet p- 40 and NaCl with proteinase K. I'm getting a very low amount of DNA and sometimes nothing ...so can somebody help me?
Hi here is a paper that might help
https://www.researchgate.net/publication/51075384_Description_of_microsatellite_markers_and_genotyping_performances_using_feathers_and_buccal_swabs_for_the_Ivory_gull_%28Pagophila_eburnea%29?ev=prf_pub
Best,
Olivier
Article Description of microsatellite markers and genotyping perform...
And amount of DNA in swabs in general can be low, you should consider this too.
well, There are many methods regarding DNA extraction, it depends which one you require and for which purpose. I think you use liquid nitrogen crass model. Use Legs for extraction.
Hi, you could try the methodology we used in :
Martín-Gálvez, D., et al. (2011). "DNA sampling from eggshell swabbing is widely applicable in wild bird populations as demonstrated in 23 species." Molecular Ecology Resources 11: 481-493.
which is based on:
Martín-Platero, A. M., et al. (2010). "Chelex-based DNA isolation procedure for the identification of microbial communities of eggshell surfaces." Analytical Biochemistry 397: 253-255.
Best,
David
It depends on what you want to amplify. The Earth Microbiome Project has good protocols for 16s metagenomics
http://www.earthmicrobiome.org/
Consider also soaking the swabs in the buffer and vortex frequently. I work with dry swabs and always let them soak overnight in the buffer then vortex for 2x 30 mins.
- Badges
- Science method