| Autor: |
McIntosh SL; Department of Chemistry, P. M. Gross Chemical Laboratory, Duke University, Durham, North Carolina 27708, USA., Nunnally BK, Nesbit AR, Deligeorgiev TG, Gadjev NI, McGown LB |
| Jazyk: |
angličtina |
| Zdroj: |
Analytical chemistry [Anal Chem] 2000 Nov 01; Vol. 72 (21), pp. 5444-9. |
| DOI: |
10.1021/ac000235t |
| Abstrakt: |
This paper describes the first use of frequency-domain fluorescence lifetime for multiplex detection of DNA restriction fragments in capillary electrophoresis (CE). The fragments were labeled with monomeric intercalating dyes that can be excited by either the 488- or 514-nm line of an argon ion laser and have lifetimes in the range of 0.5-2.5 ns. We were able to achieve multiplex lifetime detection in the CE separation of a restriction fragment digest and a DNA size ladder in the same run, for fragments shorter than 700 bp. Different gel buffer systems, including a modified polyacrylamide gel and several tris-borate-EDTA/hydroxyethylcellulose (TBE/HEC) gels, were investigated for separation and detection of the dye-labeled DNA fragments. Best results for both electrophoretic resolution and lifetime detection were obtained using a gel containing 1% high molecular weight (90,000-105,000) HEC and 0.3% low molecular weight (24,000-27,000) HEC in TBE buffer. |
| Databáze: |
MEDLINE |
| Externí odkaz: |
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