Ribeiro E A, Larcom L L, Miller D P
Department of Physics, Clemson University, South Carolina 29634-1911.
Anal Biochem. 1989 Sep;181(2):197-208. doi: 10.1016/0003-2697(89)90229-7.
Techniques for analyzing DNA distributions on agarose gels are examined by both two-dimensional and one-dimensional methods. It is demonstrated that very large errors in DNA concentration occur in such analyses unless (i) the electrophoresis is performed in a careful, reproducible manner, (ii) the films are calibrated with an internal standard, (iii) high resolution densitometry is used for analyzing the films, and (iv) appropriate background controls are used to determine the baselines for integration. Two-dimensional scanning produces more accurate results than one-dimensional scanning, but in cases where the bands are relatively uniform, the one-dimensional analysis gives good results. A technique for determining accurate distributions is described.
通过二维和一维方法研究了分析琼脂糖凝胶上DNA分布的技术。结果表明，除非满足以下条件，否则此类分析中DNA浓度会出现非常大的误差：（i）以仔细、可重复的方式进行电泳；（ii）用内标对胶片进行校准；（iii）使用高分辨率光密度计分析胶片；（iv）使用适当的背景对照来确定积分基线。二维扫描比一维扫描产生更准确的结果，但在条带相对均匀的情况下，一维分析也能给出较好的结果。本文描述了一种确定准确分布的技术。
