Southern Blot Hybridization of Digoxigenin-Labeled RNA Minisatellite Probes and Color Detection
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The following procedure describes the hybridization of digoxigenin-labeled RNA minisatellite probes ( see chapter 12 ) to Southern blots of different species and visualization of the DNA banding patterns by color detection. The method can be applied to both multi- and single-locus probes ( see Figs. 1 and 2 , respectively) and also for blots that have been screened several times with both radioactively labeled or digoxigenin-labeled probes ( 1 ).
Fig. 1. Examples of DNA fingerprints in three different species using digoxigenin-labeled RNA minisatellite 33.15 and color detection: ( a ) in man, ( b ) in the common marmoset ( Callithrix jacchus ), and ( c ) in the Waldrapp ibis ( Geronticus eremita ). F, father; M, mother; Dl and D2, their daughters; Ul, unrelated female; U2, unrelated male; 1 to 6, six related individuals from a captive population. 1.5- to 2 �g DNA digests were loaded per individual. The blots had been hybridized before to at least two other probes.
Fig. 2. DNA profiles in eight domestic pigs (1–8) and two wild boars (9 and 10) using digoxigenin labeling and color detection of a porcine minisatellite isolated from a genomic pig library, a to d indicate the alleles detected at this locus. Each lane contains approx 2 �g of digested DNA.
