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1
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QIAamp Viral RNA Mini kit (QIAGEN, Valencia, CA, USA)
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Puri?cation of viral nucleic acids (DNA and RNA viruses) from clarified stool suspension.
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32785066
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2
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QIAcube automated system (QIAGEN, Valencia, CA, USA)
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Automatic nucleic acid extraction.
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3
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AVE elution buer
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Used to elute the extracted nucleic acids.
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4
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TaqMan-based qPCR protocol
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Detection and quanti?cation of HBoV.
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5
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Applied Biosystems 7500 Real-Time PCR System and TaqMan Universal Master Mix (Applied Biosystems, Foster City, CA, USA)
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Real-time qPCR reaction system and master mix.
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6
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Primers and probe labeled with 5′-FAM and 3′-MGB
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Designed for the detection of HBoV from a conserved region of the genome.
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7
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gBlock Gene Fragment (Integrated DNA Technologies, Coralville, IA, USA)
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Preparation of a standard curve for quantifying HBoV viral load.
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8
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Primers AK-VP-F1/R1 and AK-VP-F2/R2
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Used in nested PCR reactions targeting the VP1/2 region for sequencing HBoV.
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9
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QIAquick PCR Purication Kit (QIAGEN, Valencia, CA, USA)
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Purification of the PCR amplicons obtained from nested PCR.
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10
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BigDyeTerminator v. 3.1 Cycle Sequencing Kit (Applied Biosystems, Foster City, CA, USA)
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Used for performing the sequencing reactions.
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11
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Fecal suspensions (10% w/v) prepared with Tris-calcium buer (pH = 7.2)
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Preparation of stool samples before nucleic acid extraction.
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12
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Negative control (RNAse/DNAse-free water)
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Control reagent used in each extraction procedure to ensure accuracy.
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13
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Primers and probes for TaqMan-based qPCR reactions targeting RVA, HAdV, norovirus GI and GII, sapovirus, and astrovirus
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Detection and quantification of other major gastroenteric viruses to investigate viral co-infections.
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14
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Standard curves generated from ten-fold serially diluted gBlocks dsDNA fragments
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Determining the viral loads of RVA, HAdV, norovirus GI and GII, sapovirus, and astrovirus.
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