sábado, 29 de noviembre de 2014

Genome Biology | Abstract | Enhanced methods for unbiased deep sequencing of Lassa and Ebola RNA viruses from clinical and biological samples

Genome Biology | Abstract | Enhanced methods for unbiased deep sequencing of Lassa and Ebola RNA viruses from clinical and biological samples



Enhanced methods for unbiased deep sequencing of Lassa and Ebola RNA viruses in clinical and biological samples

Christian B MatrangaKristian G AndersenSarah WinnickiMichele BusbyAdrianne D GladdenRyan TewheyMatthew StremlauAaron BerlinStephen K GireEleina England,Lina M MosesTarjei S MikkelsenIkponmwosa OdiaPhilomena E EhianeOnikepe Folarin,Augustine GobaS.Humarr KhanDonald S GrantAnna HonkoLisa HensleyChristian HappiRobert F GarryChristine M MalboeufBruce W BirrenAndreas GnirkeJoshua Z Levin and Pardis C Sabeti
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Genome Biology 2014, 15:519  doi:10.1186/s13059-014-0519-7
Published: 18 November 2014

Abstract (provisional)

We have developed a robust RNA sequencing method for generating complete de novo assemblies with intra-host variant calls of Lassa and Ebola virus genomes in clinical and biological samples. Our method uses targeted RNase H-based digestion to remove contaminating poly(rA) carrier and ribosomal RNA. This depletion step improves both the quality of data and quantity of informative reads in unbiased total RNA sequencing libraries. We have also developed a hybrid-selection protocol to further enrich the viral content of sequencing libraries. These protocols have enabled rapid deep sequencing of both Lassa and Ebola virus and are broadly applicable to other viral genomics studies.


The complete article is available as a provisional PDF. The fully formatted PDF and HTML versions are in production.

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