METHOTREXATE-LEUCOVORIN SYNCHRONIZATION OF HUMAN-LYMPHOCYTE CULTURES - INDUCTION OF HIGH-RESOLUTION R-BANDING AND G-BANDING

  • 1 January 1984
    • journal article
    • research article
    • Vol. 4  (6) , 357-360
Abstract
Simple techniques for induction of high resolution R- and G-banding was described. A methotrexate induced S-phase block was released after 18 h by exposure to leucovorin (LV). G-bands were induced by adding ethidium bromide to the cultures followed by a modified trypsin treatment of the slides, including inhibition of trypsin activity by trypsin inhibitor. To obtain R-banded preparations 5-bromodeoxyuridine and Hoechst 33258 were simultaneously added to the LV released cultures. The synchronization technique and the R- and G-banding procedures are simpler than most previously published techniques and ensure high metaphase yield, reproducibility and slides with well-defined high resolution R- or G-banding.