1
Clinical patient material (nasopharyngeal washes from three children)
Source of HBoV-positive material for virus inoculation and replication study.
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2
HTEpC (human trachea epithelial primary cells)
Cell culture for forming pseudostratified human airway epithelium.
3
Bronchial/tracheal epithelial cell serum-free growth medium
Growth medium for maintaining HTEpC cultures.
4
Penicillin-streptomycin
Antibiotic supplementation for growth medium.
5
TrypLE Express enzyme (Invitrogen)
Enzyme for dissociation of HTEpC cells when reaching confluence.
6
Air-liquid interface medium
Medium for culturing HTEpC cells in air-liquid interface condition.
7
LHC basal medium (Invitrogen) and Dulbecco’s modified Eagle’s medium (Invitrogen)
Components of the air-liquid interface medium.
8
Type IV collagen (Sigma)
Coating agent for 12-well ThinCerts to promote cell adhesion.
9
12-well ThinCerts with 0.4-μm pore size (Greiner Bio-One)
Support for cell cultures to form pseudostratified human airway epithelium.
10
Hanks’ balanced salt solution (HBSS) (Invitrogen)
Washing reagent for the apical surface of cell cultures.
11
HBSS
Dilution buffer for clinical samples before inoculation.
12
TRIzol reagent (Invitrogen)
Reagent for RNA extraction from cells for HBoV mRNA analysis.
13
L6 lysis buffer
Buffer for viral DNA quantification.
14
Boom method reagents
For total nucleic acid isolation.
15
DNase I (Ambion)
Enzyme for DNase protection assay to distinguish between protected viral DNA and naked DNA.
16
Naked plasmid DNA containing phocid herpesvirus-1 DNA
Control DNA in the DNase protection assay.
17
Cell culture supernatant of human adenovirus type 5
Control for DNase protection assay.
18
BigDye Terminator version 1.1 cycle sequencing kit (Applied Biosystems)
Reagent kit for Sanger sequencing.
19
GeneAmp PCR system 9700 thermal cycler (Perkin Elmer)
Thermal cycler used for DNA amplification in sequencing.
20
3100 genetic analyzer (Applied Biosystems)
Instrument for electrophoresis and data collection.
21
5' RACE system, version 2.0 (Invitrogen)
Kit for rapid amplification of cDNA ends to determine 5' sequences of mRNA transcripts.
22
3' RACE system (Invitrogen)
Kit for determining 3' ends of HBoV mRNA transcripts.
23
NucleoSpin Extract II kit (Machery-Nagel)
Kit for purifying PCR products after agarose electrophoresis.
24
pCR2.1-TOPO TA vector (Invitrogen)
Vector for cloning PCR products for transformation into E. coli.
25
TOP10 Escherichia coli (Invitrogen)
E. coli strain for transformation in TOPO TA cloning.
26
T7 and M13RP primers
Primers for colony PCR to verify cloned products.
