Endoglucanase, its coding gene cel5a-h31 and its application
An endoglucanase and gene technology, applied in the field of genetic engineering, can solve problems such as slow process, achieve the effects of short production cycle, strong pH tolerance, and ensure amplification efficiency
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[0037] 1. Extraction of total RNA from rumen contents of Hu sheep
[0038] Total RNA was extracted from rumen content samples frozen at -80°C (the following lysate RL, adsorption column RA, deproteinized solution RE, rinse solution RW, and RNase Free Water were all purchased from Adelaide Biotechnology Co., Ltd.) in liquid nitrogen Grind quickly into powder, add 1ml of lysate RL for every 50-100mg sample, homogenate, shake and mix vigorously to lyse the cells. Incubate at 15-30°C for 5 minutes to completely disassemble the ribosomes. Centrifuge at 12000rpm at 4°C for 10min to remove insoluble matter. Transfer the supernatant to a clean centrifuge tube, add 0.2ml chloroform for every 1ml lysate RL, shake vigorously for 15s, centrifuge at 12000rpm, 4°C for 10min. Take the upper colorless water phase and transfer it to a new tube, add absolute ethanol half the volume of the water phase, mix well and add to the adsorption column RA, centrifuge at 12000rpm for 45s at room tempera...
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