Corley-Smith G E, Lim C J, Kalmar G B, Brandhorst B P
Institute of Molecular Biology and Biochemistry, Simon Fraser University, Burnaby, BC, Canada.
Biotechniques. 1997 Apr;22(4):690-2, 694, 696 passim. doi: 10.2144/97224st04.
A method is presented for the analysis of fluorescently labeled random amplified polymorphic DNA (FRAPD) fragments. A DNA sequencer and collection and analysis software were used to estimate the sizes of DNA fragments based on their mobilities relative to in-lane size markers. This allowed confident identification and comparison of FRAPD markers both within and between polyacrylamide gels. In comparison with analysis of RAPD products using ethidium bromide-stained agarose gels, fluorescent analysis improved the sensitivity, resolution and precision of sizing of RAPD products of about 50-2100 bp. FRAPD fragments produced from amplification of zebrafish DNA are informative as genetic markers that segregate with Mendelian inheritance. FRAPD analysis was found to be very efficient for identifying new DNA polymorphisms.
本文介绍了一种用于分析荧光标记的随机扩增多态性DNA（FRAPD）片段的方法。使用DNA测序仪以及收集和分析软件，根据DNA片段相对于泳道内大小标记的迁移率来估计其大小。这使得在聚丙烯酰胺凝胶内部和之间都能够可靠地鉴定和比较FRAPD标记。与使用溴化乙锭染色的琼脂糖凝胶分析RAPD产物相比，荧光分析提高了大小约为50 - 2100 bp的RAPD产物大小测定的灵敏度、分辨率和精确度。从斑马鱼DNA扩增产生的FRAPD片段作为与孟德尔遗传分离的遗传标记具有信息价值。发现FRAPD分析在鉴定新的DNA多态性方面非常有效。
