Author: Kolodziejek, Jolanta; Seidel, Bernhard; Jungbauer, Christof; Dimmel, Katharina; Kolodziejek, Michael; Rudolf, Ivo; Hubálek, Zdenek; Allerberger, Franz; Nowotny, Norbert
Title: West Nile Virus Positive Blood Donation and Subsequent Entomological Investigation, Austria, 2014 Document date: 2015_5_11
ID: 0vm2hhdr_21
Snippet: Step RT-PCR Kit (Qiagen, USA) following the manufacturer's instructions. Prior to sequencing, all specific amplification products were purified using PCR Kleen Spin Columns (BIO-RAD, Hercules, USA) following the manufacturer's protocol. The purified PCR fragments were then premixed with the corresponding individual PCR primers (concentration of 2 μM each) in a volume of 15 μl. Sequencing in both directions was performed by Microsynth (http://ww.....
Document: Step RT-PCR Kit (Qiagen, USA) following the manufacturer's instructions. Prior to sequencing, all specific amplification products were purified using PCR Kleen Spin Columns (BIO-RAD, Hercules, USA) following the manufacturer's protocol. The purified PCR fragments were then premixed with the corresponding individual PCR primers (concentration of 2 μM each) in a volume of 15 μl. Sequencing in both directions was performed by Microsynth (http://www.microsynth.ch/). The obtained WNV sequences were manually verified and compiled to continuous sequences. Thereafter nucleotide sequences of the new WNVs were submitted to BLAST (http://www.ncbi.nlm.nih.gov/BLAST/ ) for further comparison with other WNV sequences deposited in GenBank databases. All complete genomic WNV sequences were downloaded individually in FASTA format. Sequences determined in this study were then compared to each other and to sequences from GenBank by using the Align Plus 4 program (Scientific & Education Software). Their nucleotide identities were determined.
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