%0 Journal Article
%T Efficient recovery of whole blood RNA - a comparison of commercial RNA extraction protocols for high-throughput applications in wildlife species
%A Doreen Schwochow
%A Laurel EK Serieys
%A Robert K Wayne
%A Olaf Thalmann
%J BMC Biotechnology
%D 2012
%I BioMed Central
%R 10.1186/1472-6750-12-33
%X Despite the use of minute starting volumes, all methods produced quantifiable RNA extracts (1.4 ¨C 18.4£¿¦Ìg) with varying integrity (RIN 4.6 - 7.7), the latter being significantly affected by the storage and extraction method used. We observed a significant overall effect of the extraction method on DNA contamination. One particular extraction method, the LeukoLOCK£¿ filter system, yielded high RNA integrity along with low DNA contamination and efficient depletion of hemoglobin transcripts highly abundant in whole blood. In a proof of concept sequencing experiment, we found globin RNA transcripts to occupy up to £¿ of all sequencing reads if libraries were not depleted of hemoglobin prior to sequencing.By carefully choosing the appropriate RNA extraction method, whole blood can become a valuable source for high-throughput applications like expression arrays or transcriptome sequencing from natural populations. Additionally, candidate genes showing signs of selection could subsequently be genotyped in large population samples using whole blood as a source for RNA without harming individuals from rare or endangered species.
%K Whole blood
%K RNA preservation
%K RNA extraction
%K DNA contamination
%K Transcriptome sequencing
%K Globin transcripts
%U http://www.biomedcentral.com/1472-6750/12/33/abstract