![]() At present, I'm using Fathead minnow (FHM) cells to culture these viruses. As part of my analysis I'm conducting yield studies to determine the number of competent virions produced at various time points within the infection cycle. I'm currently working on the structural analysis of a few different Ranavirus species. Could there be some clue or advise/protocol to use could some expert render technical assistance here? Note: mutation occur greatly in We thought of performing negative staining and direct RNA sequencing, however, the tedious nature of sucrose gradient, effect of ultracentrifuge speed on the virus envelope protein, lack of specific/random primers, cost of next-gene sequence and chances of having more of host associated mRNA in nextgene sequencing data especially if the sample is not purified are challenges to us. Samples were tested negative by Haemagglutination, negative for Newcastle disease virus using PCR, negative for mycoplasma gallisepticum using PCR and negative for Avian influenza virus using PCR. All effort to amplify avian coronavirus using different sets of universally designed primers yielded either unspecific band or no detectable band at all. ![]() There was evidence of virus growth in cell culture with CPE characterized by cell rounding, syncitial cells formation, and cells detachment within 4-5day pi and within 2-3 days pi. ![]()
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