TALEN Design for Targeting kif6 Exon

Two TALENs (left and right) were designed to target the second exon of kif6. These sites flanked a BccI restriction site that was used for mutation screening. TALENs were generated using the Golden Gate method as previously described (Cermak et al., 2011 (link)). Plasmids were acquired from the Golden Gate TALEN and TAL Effector Kit (AddGene) and RVD repeat arrays were cloned into pCS2TAL3DD and pCS2TAL3RR (AddGene). Plasmids were transformed into DH5a competent cells (Invitrogen) and isolated using the QIAprep Spin Miniprep Kit (Qiagen). The following sequence targets and RVD sites were used to generate kif6 TALENs: Left target DNA TCCCTCTTATTGTTG, LeftRVDsequence NGHDHDHDNGHD NGNGNINGNGNNNGNGNN Right target DNA GCATCTCTGGGAACC RightRVDsequence NNNNNGNGHDHDHDNINN NINNNINGNNHD