A simple and efficient method for the separation and detection of small DNA fragments by electrophoresis in formamide containing agarose gels and Southern blotting to DBM-paper.
We show in this report that DNA fragments smaller than 300 bp are separated with high resolution by electrophoresis in concentrated (up to 7%) agarose gels containing 50% formamide. The separated DNA fragments can subsequently be quantitatively transferred to DBM-paper [Alwine, J.C. et al., Proc. Natl. Acad. Sci. USA (1977) 74, 5350-54] using the Southern technique [Southern, E.M., J. Mol. Biol. (1975) 98, 503-517], while preserving the sharpness of the original gel pattern. Since thin (0.2-0.4 mm) and thick (up to 5 mm) agarose slab gels can be easily handled in vertical or horizontal apparatus, this method should prove to be a very useful extention of the Southern technique, applicable to a variety of analytical and preparative purposes.
D001482:Base Composition; D004247:DNA / Q000302:isolation & purification*; D003979:Diazonium Compounds; D004587:Electrophoresis, Agar Gel / Q000379:methods; D005559:Formamides; D007202:Indicators and Reagents; D008970:Molecular Weight; D009693:Nucleic Acid Hybridization; D009838:Oligodeoxyribonucleotides / Q000302:isolation & purification*; D009841:Oligonucleotides / Q000302:isolation & purification*; D012685:Sepharose
D003979:Diazonium Compounds; D005559:Formamides; D007202:Indicators and Reagents; D009838:Oligodeoxyribonucleotides; D009841:Oligonucleotides; C015794:diazobenzyloxymethyl groups; C031066:formamide; D004247:DNA; D012685:Sepharose