A kind of hippocampal oxytocic agent and its coding gene
An amino acid, sequence technology, applied in the fields of biochemistry and molecular biology
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Embodiment 1
[0019] 1. Extraction of total RNA from the whole brain of the striated hippocampus
[0020] A sexually mature linear hippocampus (Hippocampus erectus), with a body length of 5-8 cm and a body weight of about 8-10 g, was anesthetized with 20 mg / L MS222 (ethyl m-aminobenzoate methanesulfonate) for about 2 minutes, then sampling was started, and the samples were separated. The whole brain tissue was taken out, quickly frozen in liquid nitrogen, and stored in a -80°C refrigerator for later use. The total RNA of the whole brain of the striated hippocampus was extracted by Trizol reagent method, and its OD 260 / 280 = 1.90.
[0021] 2. Cloning of kiss2 cDNA
[0022] 2.1. Synthesis of the first strand of cDNA
[0023] 10 μg of total RNA samples from the whole brain of the lined hippocampus were treated with DNAase I to remove the contamination of genomic DNA, mixed with RNA Oligo dT, and subjected to reverse transcription PCR. The first strand of cDNA obtained was stored at -20°C fo...
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