与通过脉冲场凝胶电泳分离的基因组DNA进行定量杂交。
Quantitative hybridization to genomic DNA fractionated by pulsed-field gel electrophoresis.
作者信息
Leach T J, Glaser R L
机构信息
Laboratory of Developmental Genetics, Wadsworth Center, New York State Department of Health, Albany, NY, USA.
出版信息
Nucleic Acids Res. 1998 Oct 15;26(20):4787-9. doi: 10.1093/nar/26.20.4787.
Hybridization to genomic DNA fractionated by CHEF electrophoresis can vary >100-fold if the DNA is acid depurinated prior to Southern blotting. The level of hybridization is high or low depending on whether the molecule being analyzed migrates at a size coincident with or different from the size of the majority of genomic DNA in the sample, respectively. Techniques that avoid acid depurination including in-gel hybridizations and UV irradiation of DNA prior to blotting provide more accurate quantitative results. CHEF analysis of DNA molecules containing repetitive satellite sequences is particularly prone to this effect.
如果在Southern印迹之前对DNA进行酸脱嘌呤处理，与通过CHEF电泳分离的基因组DNA的杂交可能会有超过100倍的差异。杂交水平的高低分别取决于被分析的分子迁移的大小与样品中大多数基因组DNA的大小是否一致。避免酸脱嘌呤的技术，包括凝胶内杂交和印迹前对DNA进行紫外线照射，可提供更准确的定量结果。对含有重复卫星序列的DNA分子进行CHEF分析特别容易受到这种影响。
Zotero 插件