TY - JOUR
T1 - Unidirectional pulsed-field electrophoresis of single- and double-stranded DNA in agarose gels
T2 - Analytical expressions relating mobility and molecular length and their application in the measurement of strand breaks
AU - Sutherland, John Clark
AU - Monteleone, Denise C.
AU - Mugavero, Jo Ann H.
AU - Trunk, John
N1 - Funding Information:
’ This investigation was supported by the Office of Health and Environmental Research, U.S. Department of Energy, and by a grant from the National Institutes of Health (GM 35662). 2 To whom correspondence should be addressed.
PY - 1987/5/1
Y1 - 1987/5/1
N2 - Unidirectional pulsed-field electrophoresis improves the separation of single-stranded DNA molecules longer than 20 kilobases (kb) in alkaline agarose gels compared to static-field electrophoresis. The greatest improvement in separation is for molecules longer than 100 kb. The improved resolution of long molecules with unidirectional pulsed-field electrophoresis makes possible the measurement of lower frequencies of single-strand breaks. The analytical function that relates the length and mobility of single-stranded DNA electrophoresed with a static field also applies to unidirectional pulsed field separations. Thus, the computer programs used to measure single-strand breaks are applicable to both undirectional pulsed- and static-field separations. Unidirectional pulsed-field electrophoresis also improves the separation of double-stranded DNA in neutral agarose gels. The function relating molecular length and mobility for double-stranded DNA separated by unidirectional pulsed-field electrophoresis is a superset of the function for single-stranded DNA. The coefficients of this function can be determined by iterative procedures.
AB - Unidirectional pulsed-field electrophoresis improves the separation of single-stranded DNA molecules longer than 20 kilobases (kb) in alkaline agarose gels compared to static-field electrophoresis. The greatest improvement in separation is for molecules longer than 100 kb. The improved resolution of long molecules with unidirectional pulsed-field electrophoresis makes possible the measurement of lower frequencies of single-strand breaks. The analytical function that relates the length and mobility of single-stranded DNA electrophoresed with a static field also applies to unidirectional pulsed field separations. Thus, the computer programs used to measure single-strand breaks are applicable to both undirectional pulsed- and static-field separations. Unidirectional pulsed-field electrophoresis also improves the separation of double-stranded DNA in neutral agarose gels. The function relating molecular length and mobility for double-stranded DNA separated by unidirectional pulsed-field electrophoresis is a superset of the function for single-stranded DNA. The coefficients of this function can be determined by iterative procedures.
KW - DNA damage
KW - DNA mobility
KW - agarose gel electrophoresis
KW - pulsed-field electrophoresis
KW - single-strand breaks
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U2 - 10.1016/0003-2697(87)90427-1
DO - 10.1016/0003-2697(87)90427-1
M3 - Article
C2 - 3605613
AN - SCOPUS:0023193618
SN - 0003-2697
VL - 162
SP - 511
EP - 520
JO - Analytical Biochemistry
JF - Analytical Biochemistry
IS - 2
ER -