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DNA vectors Expression vector for AHCY was obtained by amplifying the mouse AHCY ORF from the HA-AHCY vector 59 using the primers 5 -ATAGTCGACGCCACCATGTACCCATACGATGTT-3 and 5 -TATGCTAGCTTCAGTAGCGGTAGTGATCAGGCT-3 , then ligating the SalI/NheI-digested PCR product into the SalI/NheI-digested pSELECT-HYGRO-MCS vector (Invivogen) using Ligation high Ver.2 (Toyobo)

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Insider Tip: Peptide needles are typically quite thin and so cause much less discomfort than a shot at the doctors office

The tibial nerve was transected at 2 mm proximal to the sutures at which the first tracing was performed, and the proximal tibial nerve stump was soaked into 5% FG in saline for 60 min with Parafilm TM protection and the excessive tracer removed as described above