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. 1997 Aug 15;326(Pt 1):131–138. doi: 10.1042/bj3260131

Purification and staining of intact yeast DNA chromosomes and real-time observation of their migration during gel electrophoresis.

S Gurrieri 1, C Bustamante 1
PMCID: PMC1218646  PMID: 9337860

Abstract

In the past few years, fluorescence microscopy has been used successfully to characterize the motion of intermediate-size DNA molecules (50-500 kbp) during steady- and pulsed-field gel electrophoresis. However, experimental difficulties had prevented the application of this technique to the direct observation of longer DNA chromosomes (1-2 Mbp). In the present study a particular procedure was followed for the purification and staining of chromosomal yeast DNA to protect it from shear forces. Also, a new highly fluorescent DNA-labelling dye, YOYO-1, was employed to improve brightness and contrast. Finally, the motion of such long DNA molecules (1-2 Mbp) was characterized under steady-field electrophoresis conditions. An accurate description of the molecular mechanisms of motion of such long molecules should provide the basis for a detailed analysis of the mechanisms responsible for DNA trapping.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

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