Hello everyone, I am new in the field of PCR-RFLP and I am planning to use it for clustering yeast isolates. I am using HaeIII, HhaI and HinfIII to digest the ITS region and using GelAnalyzer software for the analysis of the band sizes. I am getting similar restriction patterns while repeating the procedure on the same isolates. However, the band sizes have been shown to be nearly 70-80 bp higher in the second run, in the software. I doubt, this might be because of the lower resolution of the bands. Please suggest, how can I obtain accurate band sizes in each gel? Moreover, in most of the papers I have read, I have observed that, the authors depict the band sizes in multiples of 5, like 355bp, 360bp etc. Why is it so?
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Also check https://link.springer.com/article/10.1007/s10811-018-1491-5
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