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Electrophoresis and movements of fluorescence pattern after photobleaching of large DNA fragments in agarose gels
Author(s) -
Wu Chi,
Wang Zhulun,
Chu Benjamin
Publication year - 1990
Publication title -
biopolymers
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.556
H-Index - 125
eISSN - 1097-0282
pISSN - 0006-3525
DOI - 10.1002/bip.360290304
Subject(s) - chemistry , photobleaching , agarose , dna , fluorescence , gel electrophoresis of nucleic acids , fluorescence recovery after photobleaching , electrophoresis , biophysics , agarose gel electrophoresis , gel electrophoresis , chromatography , biochemistry , optics , physics , membrane , biology
Abstract By combining electrophoresis with movements of fluorescence pattern after photobleaching (MOFPAP), which is abbreviated as EMOFPAP, we are able to measure electrophoretic mobilities of large DNA fragments in an agarose gel within a fairly short time scale (about 10 min or even down to 1 min). The new method represents a significant improvement in experiment time when compared with the time (typically on the order of hours) required to determine the average electrophoretic mobility of large DNA fragments in agarose gels by means of either conventional gel electrophoresis or pulsed‐field gel electrophoresis. In this article, we present the EMOFPAP experimental setup and consider optical conditions, including beam profile geometry and fluorescence pattern formation. A realistic formula that can explain the parameters governing the EMOFPAP method using our present optical setup has been derived. A comparison of results between experimental and computer simulation data is made, and an optimization of the EMOFPAP method is proposed.