I'm interested in amplifying small quantities of genomic DNA (~2 ng/μl) for use in RadSequencing. The specimens include both fresh and 10+ year old sand flies (Lutzomyia) stored in ethanol at 4C.
Does anyone have any recommendations regarding WGA MDA kits? I'm aware of kits offered by Qiagen and Sygnis, which both utilize Phi29 polymerase, but vary significantly in price.
Any input would be greatly appreciated!
I know of both positive and negative experiences when using WGA material for RAD-Seq. If you go that route, I would include a check of the complexity of the resulting amplified material... you might get a lot of DNA but lose heterozygosity at loci. If you want to skip that step, you might want to consider nextRAD, which has been done on psyllid, thrips, mosquitos, etc with limited input. See http://biorxiv.org/content/early/2016/04/06/046540 for example.
Send an email to me at [email protected] if you would be interested in outsourcing this. (Conflict of interest note: I work at SNPsaurus).
Good luck with your genotyping!
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