Grass carp thymosin extrasin beta11 gene sequence
A thymosin and grass carp technology, applied in genetic engineering, plant genetic improvement, hormone peptides, etc.
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[0020] The present invention will be further elaborated below through the following specific embodiments, but the content of the present invention is not limited thereto at all.
[0021] 1. Extraction of total RNA
[0022] Select the experimental material—healthy female grass carp, body length 570mm, about 1.5 winter age (18 months), 2.4kg. Fresh grass carp intestinal tissues were taken out, quickly frozen in liquid nitrogen and stored in a -80°C refrigerator for later use. Total RNA was extracted by one-step method using TRIzol (Invitrogen Corporation), and the quality of total RNA was detected by formaldehyde denaturing gel electrophoresis and DNA / RNA calculator (Qenequant).
[0023] 2. Synthesis of the first strand of cDNA
[0024] Mix 5ug total RNA of grass carp intestinal cells with 1ul (12uM) of reverse transcription primers (5-terminal core primer and linker primer), heat at 70°C for 2 minutes, place on ice immediately, then add 5x buffer, 10mM dNTP mixture, 20mM DTT...
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