Rapid transfer of DNA from agarose gels to nylon membranes

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ID: 291320
1985
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Abstract
The unique properties of nylon membranes allow for dramatic improvement in the capillary transfer of DNA restriction fragments from agarose gels (Southern blotting). By using 0.4 M NaOH as the transfer solvent following a short pre-treatment of the gel in acid, DNA is depurinated during transfer. Fragments of all sizes are eluted and retained quantitatively by the membrane; furthermore, the alkaline solvent induces covalent fixation of DNA to the membrane. The saving in time and materials afforded by this simple modification is accompanied by a marked improvement in resolution and a ten-fold increase in sensitivity of subsequent hybridization analyses. In addition, we have found that nylon membrane completely retains native (and denatured) DNA in transfer solvents of low lonic strength (including distilled water), although quantitative elution of DNA from the gel is limited to fragments smaller than 4 Kb. This property can be utilized in the direct electrophoretic transfer of native restriction fragments from polyacrylamide gels. Exposure of DNA to ultraviolet light, either in the gel or following transfer to nylon membrane, reduces its ability to hybridize.
Reference Key
openalex_W1967999707 Use this key to autocite in the manuscript while using SciMatic Manuscript Manager or Thesis Manager
Authors Ken C. Reed, David A. Mann
Journal Nucleic Acids Research
Year 1985
DOI
10.1093/nar/13.20.7207
URL
https://doi.org/10.1093/nar/13.20.7207
Keywords Keywords not found

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