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1
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Phosphate Buffered Saline (PBS)
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Preparation of 10% fecal suspension.
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19381259
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2
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0.2 nm bacterial filter
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Filtration to remove bacterial contaminants.
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3
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Nucleases
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Digestion of non-viral nucleic acids.
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4
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RNeasy Mini Kit (QIAGEN)
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Extraction of viral nucleic acids.
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5
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Degenerate oligonucleotide primed (DOP) PCR primers
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Non-specific amplification of viral DNA.
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6
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AmpliTaq Gold (ABI)
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DNA polymerase used in PCR amplification.
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7
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M13 universal forward and reverse primers
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Primers used to amplify cloned DNA inserts.
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8
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BigDye 3 terminator chemistry (ABI)
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Sequencing reagent for Sanger sequencing.
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9
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Agencourt AMPure
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Purification of PCR products.
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10
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Agencourt CleanSEQ
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Purification of sequencing reactions.
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11
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TopoTA vector (Invitrogen)
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Cloning vector for DOP amplified products.
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12
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One Shot Topo10 competent cells (Invitrogen)
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Transformation host cells for cloning.
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13
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GeneAmp PCR Buffer (ABI)
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Buffer for PCR reactions.
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14
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MgCl2
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Magnesium chloride used in PCR reactions.
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15
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dNTPs (Finnzymes)
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Deoxynucleotide triphosphates used in PCR reactions.
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16
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Specific primers designed from DOP sequences
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To fill gaps in the HBoV2 genome and verify sequence accuracy.
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17
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Semi-DOP PCR approach primers
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To extend viral genome sequences upstream and downstream.
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18
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Nested consensus PCR primers (Adel-OF, Adel-OR, Adel-IF, Adel-IR)
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Detection and amplification of HBoV2 and HBoV from fecal samples.
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