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Hello Dr. Schwaner, I wonder if you got some helpful feedback from your previous email? My lab is working on ranavirus tests (our first try) for reptiles and having similar problems. Anything you can send along would be appreciated. Best wishes, Rachel On Mon, Dec 2, 2013 at 2:09 PM, Terry Schwaner <[log in to unmask]> wrote: > My colleagues and I loaded two gels with a 200 bp DNA ladder, a blank > (negative control), two amplified *Ranavirus* samples (positive > controls), and amplified samples of gDNA from tail tips from frogs, toads > and salamanders using MCP-1F and MCP-6R primers. The results in the > attachment were suggested the presence of *Ranavirus* for the two > positive controls but the primers did not result in amplicons for the > negative control, or for the frog and toad samples. However, bright bands > appeared in the lanes for the four newt samples (*N. viridescens*) but > the size of the bands was only about 400 bp. Upon sequencing, the newt > amplicons exhibited lots of “noise” and did not give any matches when > blasted in GenBank. Thinking the result might be linked to a low annealing > temperature (50C for 2 minutes), we raised the annealing temperature to > 55C. Oddly, the salamander bands were bright as ever and the positive > samples a bit lighter! Has anyone else experienced this? Any explanations > would be helpful! And thank you for considering the question. > > > > *Terry Schwaner, Ph.D.* > > Dean, Center for Life & Health Sciences > 1101 Sherman Drive, PH301D > Utica, NY 13501-5394 > P: (315)792-5376 > > F: (315)731-5834 > > *[log in to unmask] <[log in to unmask]>* > > > > [image: MVCC Logo2] > > > > Strategic ■ Input ■ Activator ■ Learner ■ Arranger > > > -- Dr. Rachel M. Goodman Assistant Professor of Biology Hampden-Sydney College, VAhttp://www.hsc.edu/Academics/Academic-Majors/Biology/Professors/Rachel-Goodman.html