|
1
|
Swabbing separation grids
|
Collection of influent sewage at the Makana Wastewater Treatment Plant
|
31786741
|
View Related Publication
|
|
2
|
Glycine buffer (0.25 N, pH 10) containing NP-40
|
Elution of homogenized tissue of mussel samples
|
|
3
|
0.20 μM membrane (GVS filter technology, USA)
|
Filtration of procedure supernatant
|
|
4
|
QIAamp Viral RNA Mini Kit (Qiagen, USA)
|
Extraction of viral nucleic acid from swab and mussel samples
|
|
5
|
Primers 2028 (+), 2029 (), 2030 (+), 2031 ()
|
Nested PCR assays for molecular detection of HBoV DNA
|
|
6
|
Taq DNA Polymerase 2× Master Mix RED PCR Kit (Ampliqon Bio Reagents and Molecular Diagnostics, Denmark)
|
PCR reactions for detecting HBoV DNA
|
|
7
|
Negative template controls
|
To discount the possibility of cross-contamination in PCR
|
|
8
|
pJET 1.2/blunt (CloneJET PCR Cloning Kit; Thermo Scientific, USA)
|
Cloning of PCR products prior to sequencing
|
|
9
|
MEGA 7 software
|
Determination of phylogenetic relationships using aligned nucleotide sequences
|
