New technique for high resolution DNA sizing in epi-illumination
✍ Scribed by J. M. Schins; A. Agronskaya; B. G. de Grooth; J. Greve
- Publisher
- John Wiley and Sons
- Year
- 1998
- Tongue
- English
- Weight
- 90 KB
- Volume
- 32
- Category
- Article
- ISSN
- 0196-4763
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✦ Synopsis
We present a high-resolution DNA-sizing technique based on the principles of flow cytometry, using a high numerical aperture objective and epiillumination. The new technique, designed for small fluorescing samples/particles (sub-micron diameter) suspended in a weakly fluorescent medium, makes use of an additional focus for high-precision particle localisation. This way, only those particles are considered that flow exactly through a well-defined volume. Results are presented for fluorescent beads, as well as for YOYO-stained plasmids containing 5,500 basepairs. The latter were measured with 6.2% resolution, setting a new limit to flow-based sizing of DNA.