Rprotk ro

Nphp1^gt and Cep290^fl mice were previously described [22 (link), 30 (link)] and obtained from the Jackson laboratory (Nphp1^tm1Jgg/J, #013169; Cep290^tm1Jgg/J, #013701). iCre75 mice were a generous gift from Dr. Ching-Kang Chen [31 (link)]. The lack of rd1 mutation (in Pde6b) was confirmed by PCR using primers described in [25 (link)]. The rd8 mutation (in Crb1) was eliminated by breeding. All animals used in this study were Pde6b^+/+;Crb1^+/+. For genotyping, mouse tail snips were collected at the time of weaning (P19-P24) or after euthanasia. Tail DNAs were extracted by Proteinase K digestion (Sigma-Aldrich; RPROTK-RO) in Tail Lysis Buffer (10 mM Tris pH 8.0, 100 mM NaCl, 10 mM EDTA, 1% SDS, 0.3 mg/ml Proteinase K) followed by ethanol precipitation. Genotyping was conducted by PCR using GoTaq G2 Flexi DNA polymerase (Promega) and primers listed in Table 1. PCR protocols are available upon request. All animals were maintained in 12-hour light/dark cycles and fed ad libitum standard mouse chow. All animal procedures were approved by the Institutional Animal Care and Use Committee (IACUC) of the University of Iowa (Protocol#: 8011301) and conducted following the recommendations in the Guide for the Care and Use of Laboratory Animals of the National Institutes of Health.