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1
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Millipore type HA negatively charged membrane filter
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Virus adsorption with a 0.45 mm pore size and a 142 mm diameter.
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19656966
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2
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Non-organic elution buffer [0.05 M KH2PO4, 1.0 M NaCl, 0.1% (v/v) Triton X-100, pH 9.2]
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Recovery of the adsorbed viruses.
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3
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PEG-6000
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Precipitation step in virus recovery.
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4
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QIAamp DNA Blood Mini kit (Qiagen)
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Extraction of viral nucleic acids.
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5
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188F (59-GACCTCTGTAAGTACTATTAC-39) and 542R (59-CTCTGTGTTGACTGAATACAG-39) primers
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Amplification of a 354 bp fragment of the HBoV genome for obtaining a DNA standard for HBoV quantitative PCR.
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6
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StrataClone PCR cloning kit (Stratagene)
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Sequencing and cloning the PCR fragment into the pSC-Aamp/kan vector.
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7
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Quant-iT dsDNA HS assay (Invitrogen)
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Determining the concentration of purified plasmid DNA; fluorescence measurement.
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8
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HBoV-NP1-F2421 (59-TGGCAGACAACTCATCACAG-39), HBoV-NP1-R2544(59-TCTTCGAAGCAGTGCAAGAC-39), and HBoV_Probe-2494 [59-(6FAM)ATCATCAGGAACACCC-AATCAGCCfAC(BHQ1)-39]
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Primers and Taqman probe designed for newly developed real-time PCR to quantify HBoV DNA in concentrated water samples.
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9
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Quantitect probe PCR kit (Qiagen)
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Real-time PCR conducted in a 20 ml reaction volume.
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10
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DNase/RNase-free water
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Used as a negative control during PCR.
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11
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DNase I
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Treatment of river-water samples to confirm that HBoV DNA detected is protected from DNase degradation inside intact virus particles.
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