Goodwin P M, Johnson M E, Martin J C, Ambrose W P, Marrone B L, Jett J H, Keller R A
Chemical and Laser Sciences Division, Los Alamos National Laboratory, NM 87545.
Nucleic Acids Res. 1993 Feb 25;21(4):803-6. doi: 10.1093/nar/21.4.803.
Large, fluorescently stained restriction fragments of lambda phage DNA are sized by passing individual fragments through a focused continuous wave laser beam in an ultrasensitive flow cytometer at a rate of 60 fragments per second. The size of the fluorescence burst emitted by each stained DNA fragment, as it passes through the laser beam, is measured in one millisecond. One hundred sixty four seconds of fluorescence burst data allow linear sizing of DNA with an accuracy of better than two percent over a range of 10 to 50 kbp. This corresponds to analyzing less than 1 pg of DNA. Sizing of DNA fragments by this approach is much faster, requires much less DNA, and can potentially analyze large fragments with better resolution and accuracy than with gel-based electrophoresis.
通过在超灵敏流式细胞仪中以每秒60个片段的速率使单个λ噬菌体DNA的荧光染色大片段通过聚焦连续波激光束来对其进行大小测定。每个染色的DNA片段通过激光束时发出的荧光猝发大小在一毫秒内进行测量。164秒的荧光猝发数据可实现DNA的线性大小测定，在10至50千碱基对的范围内，准确度优于2%。这相当于分析不到1皮克的DNA。用这种方法对DNA片段进行大小测定要快得多，所需的DNA少得多，并且与基于凝胶的电泳相比，有可能以更高的分辨率和准确度分析大片段。
