Pcr grade proteinase k

Genomic DNA (obtained from a 2 mm ear punch collected at post-natal day (P) 14) was extracted using DirectPCR Lysis Reagent (Tail) (Viagen Biotech, Los Angeles, CA) and PCR Grade Proteinase K (Sigma-Aldrich) according to the manufacturer’s protocol. Briefly, the ear punch sample was placed in 100 μL DirectPCR Lysis Reagent (Tail) (Viagen Biotech, Los Angeles, CA) containing 1 μL PCR Grade Proteinase K. The sample was incubated overnight at 55°C, and subsequently incubated at 85°C for 45 min. Two separate PCR reactions were needed to properly describe each individual animal: one detected the Gclc(wt) and Cre(Tg) alleles (Fig. 1A; left lane for each mouse), and the other detected the Gclc(f) allele (Fig. 1A; right lane for each mouse). Gclc alleles were determined using 5^′-CTATAATGTCCTGCACTGGG and 5^′-TAGTGAACGCTGTTAAAGG and 5′-CGGGTGTTGGGTCGTTTGT and the presence of the Cre transgene was identified using 5^′-GCGGTCTGGCAGTAAAAACTATC and 5^′-GTGAAACAGCATTGCCACTT, as previously described^{27 (link)}.