Summary
Type: TLC
Application #: 402170
Catalog#: 1122026 - Cellulose 300 PEI TLC Plates, polyester backed, 100um, 20x20cm 25/pk
Detection: Adduct spots located by autoradiography at -80°C using intensifying screens.
Conditions: TLC-Analysis: The labelling mix was applied to PEI-cellulose plates (10 x 10 cm) with a wick (10 x 7 cm) of MN no. 440 paper stapled to the top. Elution in the D1 direction was performed overnight with sodium phosphate buffer (1 M, pH 6.0), the wicks containing the bulk radioactivity (labelled non-adducted nucleotides, phosphate) were cut off and discarded, the plates were washed subsequently with water, ammonium formate (0.1 M, pH 2.5), and water and dried. Elution in D2 (in the opposite direction to D1) and D3 (perpendicular to D2, from left to right on the autoradiographs shown in the figure) were performed with buffers of different ionic strength according to the different mobilities of the adducts. D2: 3.5 M lithium formate, 8.5 M urea, pH 3.5; D3: 0.8 M lithium chloride, 0.5 M Tris/HCl, 8.5 M urea, pH 8.0. Finally, the plates were eluted with 1.7 M sodium phosphate (pH 6.0) overnight on to a wick of MN no. 615 paper; again the wicks were cut off, the plates washed with water and dried.
Authors: Brockstedt, U.
Source: Z. Lebensm. Unters. Frosch. A 207 (1998) 472-476