Use of resonance ionization spectroscopy to detect DNA bands on ultrathin spin-coated gels.
Development of alternative electrophoresis procedures are necessary for large volume sequencing and mapping studies. The use of stable isotopes as DNA labels and ultrathin gels promises to greatly increase the rate of sequencing. Spin coating is presented as an alternative method for producing ultrathin polyacrylamide gels. The technique has the potential of producing gels of micron to submicron thicknesses by varying the viscosity of the acrylamide solution and the spinning speed. Thirty micron thick 6% (weight %) gels were produced in this manner. Tin-labeled DNA oligomers were electrophoresed and detected using sputter-initiated resonance ionization spectroscopy (SIRIS). The usefulness of SIRIS and laser atomization RIS (LARIS) to sample the surface and deeper layers of 240 microns thick gels was investigated. With LARIS, whole cross-sections of the gel can be atomized, possibly allowing complete sampling of labels.
Doktycz
M J
MJ
Biology Division, Oak Ridge National Laboratory, TN 37831-8077.
Gibson
W A
WA
Arlinghaus
H F
HF
Allen
R C
RC
Jacobson
K B
KB
D004247:DNA / Q000302:isolation & purification*; D004591:Electrophoresis, Polyacrylamide Gel / Q000295:instrumentation / Q000379:methods*; D005069:Evaluation Studies as Topic; D017422:Sequence Analysis, DNA / Q000379:methods*; D013057:Spectrum Analysis / Q000379:methods*; D014001:Tin
D014001:Tin; D004247:DNA