RNA from positive samples acquired from virus culture was extracted by using a viral RNA extraction kit (QIAGEN, Valencia, California, USA), according to the manufacturer's instructions.
Nucleic acid was extracted using a viral nucleic acid extraction kit (Geneaid, Taiwan) from 200 μL of 10% fecal suspension to a final volume of 50 μL of RNase-free H20.
Swabs were placed in 1.5-mL virus transport medium; all environmental samples were transported at 4°C within 36 hours to Institut Pasteur in Cambodia for subtype H5N1 testing by real-time reverse transcription PCR (rRT-PCR) after RNA extraction by using a viral RNA kit (QIAamp, QIAGEN, Valencia, CA, USA) and for virus isolation after inoculation onto MDCK cells.
We used a viral RNA extraction kit (iNtRON Biotechnology, Seongnam, South Korea) to extract RNA from the supernatant of the tick homogenates.
Viral nucleic acids were extracted by using a MiniBest Viral RNA/DNA Extraction Kit (TaKaRa, Dalian, China).
Viral RNA was extracted from serum by using a QIAamp viral RNA extraction kit (QIAGEN, Hilden, Germany), and reverse transcription PCR (RT-PCR) was performed essentially as described elsewhere (14 ), with the exception that 140 μL was not available from every bat.
The viral RNA was extracted from 100 μL of the harvested cell culture using a QIAamp viral RNA extraction kit (Qiagen, Germany), as per the manufacturer's protocol.
Viral RNA was recovered from infected cells in the supernatant by using a QIAquick viral RNA extraction kit (QIAGEN, Valencia, CA, USA); genome amplification was performed by using a 1-step reverse transcription PCR (RT-PCR) (4) and 2 set of oligonucleotides designed to amplify the entire N gene in overlapping PCR products.
The nucleic acid from the phage ZZ1 was extracted and purified from phage lysate using a MiniBEST Viral RNA/DNA Extraction Kit Ver. 4.0 (TAKARA BIO Inc., Tokyo, Japan) according to the manufacturer's protocol.
RNA was extracted from 140 µL of cell culture supernatant by using a QIAamp viral RNA extraction kit (QIAGEN GmbH, Hilden, Germany) according to the manufacturer's recommendations.
