Maniatis T, Jeffrey A, van deSande H
Biochemistry. 1975 Aug 26;14(17):3787-94. doi: 10.1021/bi00688a010.
We describe the use of polyacrylamide gel electrophoresis to estimate chain lengths of double- and single-stranded DNA molecules in the size range 20-1000 base pairs (or nucleotides). Double-stranded DNA molecules of known length produced either by organic synthesis or by restriction endonuclease digestion of viral DNAs were used as standards. The relative electrophoretic mobilities of these standards were examined on both nondenaturing (aqueous) polyacrylamide gels and on denaturing gels containing 7 M urea or 98% formamide. Electrophoretic mobility of DNA is a linear function of the log of molecular weight if appropriate conditions are used, although exceptions are noted. Chain lengths can be conveniently estimated by using as standards bacteriophage gamma DNA restriction fragments or commercially available tracking dyes.
我们描述了使用聚丙烯酰胺凝胶电泳来估计大小在20至1000个碱基对（或核苷酸）范围内的双链和单链DNA分子的链长。通过有机合成或病毒DNA的限制性内切酶消化产生的已知长度的双链DNA分子用作标准品。在非变性（水性）聚丙烯酰胺凝胶以及含有7M尿素或98％甲酰胺的变性凝胶上检测这些标准品的相对电泳迁移率。如果使用适当的条件，DNA的电泳迁移率是分子量对数的线性函数，不过也有例外情况。通过使用噬菌体γDNA限制性片段或市售的示踪染料作为标准品，可以方便地估计链长。
