RNA extraction from lizard

[Dan_cham]

Hello all

I want to sequence the transcriptome of a new organism (species of lizard)

i order to do that, i need first to extract rna from fresh tissue. i follow protocol but I got an rna result that suggesting that my sample was degredated. but because I don't know many people who try to extract rna from reptiles i dont know if maybe the sample suppose to be different from standard extraction from mammals.

attached please find a pdf version of bio analyzer and gel electrophoresis of my samples (samples 11 and 12)

will be happy to hear ideas

Dan

[Wallysb01]

We've used the mirVana kit from Ambion on lizard tissues and had no real problems and don't see any differences in the prep with mammals. Usually RNA degradation can be avoid by just processing the samples quickly. Dissecting and getting into cold extraction/lysis buffer quickly is key. Then flash freeze for storage or proceed quickly through the rest of the protocol.

[ECO]

Uh....you attached a PDF of a Ruan RNA-seq paper...just FYI.

Flash freeze, grind, into Tri reagent of choice.