Gray J W, Dean P N, Fuscoe J C, Peters D C, Trask B J, van den Engh G J, Van Dilla M A
Biomedical Sciences Division, Lawrence Livermore National Laboratory, CA 94550.
Science. 1987 Oct 16;238(4825):323-9. doi: 10.1126/science.2443974.
Dual-beam high-speed sorting has been developed to facilitate purification of chromosomes based on DNA staining with the fluorescent dyes Hoechst 33258 and chromomycin A3. Approximately 200 chromosomes per second of two types can be sorted from a suspension of chromosomes isolated from human lymphoblasts while fluorescent objects (chromosomes, debris fragments, chromosome clumps, and nuclei) are processed at the rate of about 20,000 per second. This sorting rate is approximately ten times that possible with conventional sorters. Chromosomes of a single type can be sorted with a purity of about 90 percent. DNA from the sorted chromosomes is suitable for construction of recombinant DNA libraries and for gene mapping.
双光束高速分拣技术已被开发出来,以便基于用荧光染料赫斯特33258和嗜铬菌素A3对DNA进行染色来促进染色体的纯化。从人淋巴母细胞分离出的染色体悬浮液中,每秒大约可以分拣出两种类型的200条染色体,而荧光物体(染色体、碎片、染色体团块和细胞核)的处理速度约为每秒20000个。这种分拣速度大约是传统分拣器的十倍。单一类型的染色体可以以约90%的纯度进行分拣。分拣出的染色体中的DNA适用于构建重组DNA文库和基因图谱。