A comparative study on the effect of various detergents in human chromosome G-banding prior to tryptic digestion

Abstract

The effect of treatment of chromosomes with various detergents prior to tryptic banding was investigated. The pre-treatment improved the resolution of banding for most of the chromosomes. The effectiveness of the detergents varied greatly. The number of chromosome pairs with optimal banding found after the use of different detergents was as follows: 19 of the possible 23 for emulphogene, 15 for Nonidet, 9 for Triton X-100, 10 for Tween 40, 4 for Tween 80, 4 for DOC and 18 for SDS compared to 3 for trypsin without using detergent. Optimal banding was as defined by the Paris Conference (1971) map. The improvement of banding was shown to be due to detergent-induced changes in the chromosomal proteins. When the chromosomes were treated first with trypsin followed by the detergent no improvement of chromosomal banding was observed. The detergents showed a degree of specificity towards individual chromosomes; certain chromosomes were found to be better banded with a particular detergent. Pretreatment of chromosomes with a combination of two detergents — simultaneously or consecutively — was found not to be additive. After such treatment the chromosome bands were disrupted. Pretreatment with different detergents sometimes changed the topography of chromosome banding, i.e., the relative location of the bands from the centromere. These findings suggest that the proteins attached to the DNA of the chromosomes were removed or loosened from different sites by the various detergents. — For chromosomes 8, 9, 20 and X, additional bands not reported previously were detected.