Cressdnaviruses of Cynopterus bat

This study used metagenomic to characterize cressdnaviruses in a rectal swab sample of a GGreater short-nosed fruit bat (Cynopterus sphinx). Two aliquots of the sample in question were retrieved from the -80 degC freezer and centrifuged at 1800 x g and 4 degC, for 15 min. Eukaryotic and bacterial cell debris was removed by passing the supernatants through a filter with 0.45 um pores. The first aliquot (the untreated sample) was used for nucleic acid extracted with the RNA Easy Fast Tissue/Cell kit (TIANGEN). In the second aliquot (treated sample), the unprotected nucleic acids were digested with a cocktail of enzymes - 15 U Turbo DNase, 20 U Benzonase, 20 U RNase One, in 10x Turbo DNase buffer - and 30 mM EDTA was then added and incubated with the mixture for 30 minutes to stop the enzymatic reaction (120,121). The nucleic acids obtained from the two sample aliquots were reverse-transcribed and the libraries were prepared using NEBNext Ultra II library preparation kit, in accordance with the protocol recommended by the manufacturer (Illumina) for use with FFPE RNA. The libraries were pair end-sequenced on an Illumina Novogene 6000 device based in Nanjing, China.