I've been working with RNA isolated from cells obtained by laser
capture microdissection. Isolating these cells and subsequently its
RNA is very laborious and as the cells are in low abundance, I'm
limited in the RNA that I can obtain. We are talking picogram
quantities of total RNA per cell. Realistically, by pooling samples I
could bring the amount of total RNA up to the nanogram range.
Given the limited amount of RNA that we will have available the
standard RNA-seq protocols which require 1-10 micrograms of total RNA
is simply not reasonable.
Does anyone have any experience in preparing samples for RNA-seq from
low abundance RNA? I would appreciate any recommendations that you
have.
capture microdissection. Isolating these cells and subsequently its
RNA is very laborious and as the cells are in low abundance, I'm
limited in the RNA that I can obtain. We are talking picogram
quantities of total RNA per cell. Realistically, by pooling samples I
could bring the amount of total RNA up to the nanogram range.
Given the limited amount of RNA that we will have available the
standard RNA-seq protocols which require 1-10 micrograms of total RNA
is simply not reasonable.
Does anyone have any experience in preparing samples for RNA-seq from
low abundance RNA? I would appreciate any recommendations that you
have.
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