I am working on the DNA damage on Adineta ricciae bdelloid rotifers upon the desiccation. I'm resolving chromosomes on CHEF DR III system (BioRad). The probem is that I see the DNA damage even in wet samples. The only reference I found on the literature is about Adineta vaga from Gladyshev and Meselson (2008) paper ( http://www.pubmedcentral.nih.gov/articlerender.fcgi?artid=2278216&tool=pmcentrez&rendertype=abstract ). Do you have any experience in PFGE? The conditions I used in my runs were:
14-09[2]: 22 h at 14oC, 50-90 s switch times, 6 V/cm 120o angle in 0.5x TBE buffer
20-9[long2]: 72 h in 3 blocks:
Block 1: 14°C, 20 min switch time, 2 V/cm, 96° for 24 h in 1xTAE buffer
Block 2: 14°C, 25 min switch time, 2 V/cm, 100° for 24 h in 1xTAE buffer
Block 3: 14°C, 30 min switch time, 2 V/cm, 106° for 24 h in 1xTAE buffer
Many thanks,
Luke
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