Doktycz M J, Gibson W A, Arlinghaus H F, Allen R C, Jacobson K B
Biology Division, Oak Ridge National Laboratory, TN 37831-8077.
Appl Theor Electrophor. 1993;3(3-4):157-62.
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.
开发替代电泳程序对于大规模测序和图谱研究是必要的。使用稳定同位素作为DNA标记和超薄凝胶有望大大提高测序速度。旋涂法被提出作为制备超薄聚丙烯酰胺凝胶的替代方法。该技术有可能通过改变丙烯酰胺溶液的粘度和旋转速度来制备厚度为微米至亚微米的凝胶。用这种方法制备了30微米厚的6%（重量）凝胶。对锡标记的DNA寡聚物进行电泳，并使用溅射引发共振电离光谱法（SIRIS）进行检测。研究了SIRIS和激光雾化共振电离光谱法（LARIS）对240微米厚凝胶的表面和更深层进行采样的实用性。使用LARIS，可以对凝胶的整个横截面进行雾化，这可能允许对标记进行完全采样。
