and 2 and a commercially available system the cycling parameters for this amplification were 15 seconds at 94 u00b0 c 30 seconds at 61 u00b0 c 1 minute at 72 u00b0 c 10 cycles 15 seconds at 94 u00b0 c 30 seconds at 61 u00b0 c 80 seconds plus 20 seconds cycle elongation at 72 u00b0 c for 20 cycles followed by 10 minutes at 72 u00b0 c for a final extension the amplification product was cleaved with restriction endonucleases noti and sali and then subcloned into bluescript plasmid automated sequencing was then carried out using t3 and t7 primers it was confirmed to be a partial mage 10 cdna sequence 1400 base pairs which corresponded to a start at position 2770 at the 5 u2032 end and extending 660 base pairs beyond the 3 u2032 end of the genomic sequence reported by deplaen et al supra using the information obtained from the experiments described supra additional work was carried out to obtain a full length cdna clone for mage 10 as indicated the partial cdna clone was 1 4 kb long this fragment was subjected to digestion with restriction endonucleases and an hpai fragment corresponding to nucleotides 2770 3510 of the known gdna sequence was isolated and 32 p labeled using a random priming dna labeling kit the labeled probe was then used to screen two libraries from a melanoma cell line lb73 mel in the vectors pcdnai amp and pcep4 the hybridization was carried out on filters using 5 u00d7 ssc 5 u00d7 denhardt 39 s 0.