The PCR was carried out using Ex-Taq polymerase (TaKaRa, Shiga, Japan) from the adhering to ailments: ninety four , 60 sec; 60 , 60 sec; 72 , 60 sec for 35 cycles. With the amplification of both of those the Ex4a(+) and major WT1 [17AA(+) and 17AA(-) WT1] isoforms, primer pair of Ex4-F and Ex6-R was made use of. With the amplification of Ex4a(+) WT1 isoform, primer pair of 4a-F and Ex6-R was used.