Enhanced methods for unbiased deep sequencing  of Lassa and Ebola RNA viruses from clinical and  biological samples.
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.
Christian B
CB
Matranga
Kristian G
KG
Andersen
Sarah
S
Winnicki
Michele
M
Busby
Adrianne D
AD
Gladden
Ryan
R
Tewhey
Matthew
M
Stremlau
Aaron
A
Berlin
Stephen K
SK
Gire
Eleina
E
England
Lina M
LM
Moses
Tarjei S
TS
Mikkelsen
Ikponmwonsa
I
Odia
Philomena E
PE
Ehiane
Onikepe
O
Folarin
Augustine
A
Goba
S Humarr
SH
Kahn
Donald S
DS
Grant
Anna
A
Honko
Lisa
L
Hensley
Christian
C
Happi
Robert F
RF
Garry
Christine M
CM
Malboeuf
Bruce W
BW
Birren
Andreas
A
Gnirke
Joshua Z
JZ
Levin
Pardis C
PC
Sabeti
D029043:Ebolavirus / Q000235:genetics*; D019142:Hemorrhagic Fever, Ebola / Q000235:genetics / Q000821:virology; D059014:High-Throughput Nucleotide Sequencing / Q000379:methods*; D006801:Humans; D007835:Lassa Fever / Q000235:genetics / Q000821:virology; D007836:Lassa virus / Q000235:genetics*; D012367:RNA, Viral
D012367:RNA, Viral