doi: 10.2144/97224st01.
Northern blotting of RNA denatured in glyoxal without buffer recirculation
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- PMID: 9105618
- DOI: 10.2144/97224st01
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Northern blotting of RNA denatured in glyoxal without buffer recirculation
Biotechniques.
1997 Apr.
Free article
Abstract
A rapid and easy procedure for preparing Northern blots is described. RNA is denatured with glyoxal in the presence of ethidium bromide and glycerol, then electrophoresed through agarose in a buffer that does not require recirculation. Without any additional washes, the RNA is vacuum-blotted to a nylon membrane in NaOH, which simultaneously removes the glyoxal adducts. All of these steps plus prehybridization of the filter and addition of a digoxygenin-labeled probe can be completed in one day. Using standard procedures to wash the filters and detect the probe, the entire procedure can be completed within two days.
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