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mRNA expression analysis by real-time PCR Total RNA was isolated from the dissected tissues using the RNeasy Mini kit (Qiagen, CA, USA), followed by reverse transcription using the High Capacity RNA-to-cDNA kit (Thermo Fisher Scientific, Waltham, MA, USA) according to the manufacturer’s instructions

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Vitamin B12 does require the initial loading dose period, especially in very deficient cats, so improvements will not be immediate