DNA Extraction
Skin scabs from pathological tissues were centrifuged at 10,000 g for 10
min at +4 °C. The centrifuged scabs was used for viral genomic DNA
extraction using Blood-Animal-Plant DNA Preparation kit (Jena
Biosciences®, Germany). A total of 300 µl of lysis buffer was added to
200 µl of tissue homogenate and 2 µl RNAse A inhibitor was added and
vortexed for 30 seconds, then 8µl of proteinase K was added to digest
the tissue and placed on heating block at 60oC for 30
minutes. This was cooled for 5mins and 300 µl binding buffer was added
and centrifuged at 10,000g for 5mins. The supernatant was decanted into
a spin column and washed twice with wash buffer. Fifty microliter of
viral DNA was eluted into a new eppendorf tube, labeled and kept at
-4oC briefly before PCR.