Microprocedure for in situ nick translation of chromosomes

We have modified the procedure of in situ nick translation to shorten the autoradiographic exposure time from 1 month to 3 days and reduce the volume of nick translation solution by a factor of at least 10. The modified procedure can be carried out on individually chosen chromosome spreads. The procedure was used on chromosome spreads of three related lines of mouse mammary epithelium (+SA, -SA, CL-S1) with different degrees of tumorigenicity. We found that the autoradiographic silver grains that are observed following in situ nick translation were often placed at the apparent junction site of chromosome translocations or at the breakpoint of chromosomal pieces. We found also that silver grains were located above double minute chromosomes, which suggests that there are active genes in double minutes.