Abstract
DNA immobilized on a nylon membrane can be re-probed multiple times with different probes. Protocols typically recommend that DNA blots be stored either dry at room temperature or wet at 4 or −20°C after a probe is removed. This study shows substantial differences in the effect of these storage options on the performance of uncharged nylon membranes in subsequent hybridizations. Uncharged membranes, air-dried and stored at room temperature after probe removal, could not be successfully re-probed. However, excellent rehybridization results were obtained following probe removal when wet membranes were wrapped in plastic and stored at −20°C.
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Abbreviations
- BSA:
-
bovine serum albumen
- CTAB:
-
hexadecyltrimethylammonium bromide
- EDTA:
-
disodium ethylenediamine tetraacetate
- NaHPO4 :
-
sodium phosphate
- SDS:
-
sodium dodceyl
- SSC:
-
sodium citrate
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Keane, B., Lim, H.W. & Rogstad, S.H. Re-probing DNA blots: Wet is better than dry storage of uncharged nylon membranes after removing probes. Plant Mol Biol Rep 18, 17–21 (2000). https://doi.org/10.1007/BF02825289
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DOI: https://doi.org/10.1007/BF02825289