Whole Genome Sequencing of VHS Virus: Isolate DK-5p508
Viral RNA was extracted using the EZ1 Virus Mini Kit v2.0 and the EZ1 extraction robot   (Qiagen), and eluted in 60 L of elution buffer. Approximately 100 ng of RNA were reverse transcribed at 37C for 1 hour in a 20 L reaction containing 1 mM dNTPs, 500 ng of random primers and 200 units M-MLV Reverse Transcriptase (Promega). Double-stranded cDNA was then generated with random primers using Sequenase V2.0 DNA polymerase (Affymetrix)  , purified with the QIAquick PCR Purification Kit (Qiagen) and quantitated using QuantusTM Fluorometer (Promega). Libraries were prepared with the Nextera XT DNA Library Preparation Kit (Illumina) and checked for quality and size on a 2100 Bioanalyzer Instrument (Agilent) using the Agilent High Sensitivity DNA Kit.
SUB5476254
DK-5p508