Sajantila A, Lukka M
Department of Human Molecular Genetics, National Public Health Institute, Helsinki, Finland.
Int J Legal Med. 1993;105(6):355-9. doi: 10.1007/BF01222121.
The effect of a stacking gel, the pH and crosslinking agent concentration on the resolution and sharpness of PCR amplified VNTR alleles in a vertical discontinuous polyacrylamide gel electrophoresis system was investigated. The experiments show that the use of a low crosslinking agent concentration, a stacking gel and a wide pH difference between the gel buffer and the electrophoresis buffer at the beginning of the electrophoresis resulted in reduced band width and increasing resolution in silver-stained polyacrylamide gels. The importance of sharp DNA fragments is especially emphasized when analyzing multi-allelic DNA loci, that exhibit alleles differing from only few bp to few dozen bp in length, such as variable number of tandem repeat (VNTR) or short tandem repeat (STR) loci.
研究了在垂直不连续聚丙烯酰胺凝胶电泳系统中，堆积凝胶、pH值和交联剂浓度对PCR扩增的VNTR等位基因分辨率和清晰度的影响。实验表明，在银染聚丙烯酰胺凝胶中，使用低交联剂浓度、堆积凝胶以及在电泳开始时凝胶缓冲液和电泳缓冲液之间存在较大的pH差异，会导致条带宽度减小和分辨率提高。在分析多等位基因DNA位点时，尤其强调清晰DNA片段的重要性，这些位点的等位基因长度仅相差几个碱基对到几十个碱基对，例如可变串联重复序列（VNTR）或短串联重复序列（STR）位点。
