Hello Everyone,
I am working on mosquito genomics and trying to find a best protocol or method for RNA extraction from a single head and thorax part of an individual mosquito. The RNA will be used for NGS applications. Please suggest.
Hello, I send you two docs about the mosquito grinding process and RNA isolation from mosquito grind liquid, also I would like to notify you that the first doc is in French, but easy to understand.
Best regards
Dear Devojit,
depends on what you mean by "best". Time or cost efficient? Best quality?
Since you only work with insects (or part of one) you might wanna look at drosophila methods. These have been around for quite a while. Protocols for (5 mg!) of tissue with high glycogen may not be optimal.
I'm not too familiar with insects but mechanical force may be needed on dry ice/ nitrogen to prepare the tissue. Could be by using micropistils etc. You might then try to use Trizol/Trifast reagent for it seperates proteins, fat, carbohydrates and DNA from RNA quite well. You may wanna use some extraction buffer first also to have an adequat volume (aqueos phase) to handle. Then, precipitate RNA to concentrate and determine quality.
Good luck!
Best
Boris
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