Hi all,

I have been trying to knock out genes in Yarrowia lipolytica using CRISPRyl system, but it always appears right for initial verification by colony PCR (PCR products shorten as up/down homologous arm without CDS) in SC-leu plating medium, but after subculturing in YPD medium and extracting genomic DNA for re-PCR, the PCR products were reverted to wild type, or presents both two bands, which seems wrong（picture is as follows）.

Do you ever encounter this problem? Please provide some suggections. Thanks a lot!