Mouse model for mTORC1 (mechanistic target of rapamycin 1) pathways and method for building mouse model
A mouse model and pathway technology, applied in animal husbandry and other fields, can solve problems such as lack, and achieve the effect of simple method and good application prospects
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[0022] 1. Mice Breeding and Mating Program
[0023] 8-week-old sexually mature male mice and female mice were caged together, and the mice were numbered and separated into cages when they were 3 weeks old. At the same time, a small amount of mouse tail tissue was cut for subsequent genotyping. On the C57 strain of Raptor lox / lox mice, cross with RIP-Cre mice to obtain Raptor heterozygous mice, namely RIP-Cre Raptor lox / w mice, and through self-mating of Raptor heterozygous mice , to obtain Raptor lox / lox mice with or without RIP-Cre. The former is to specifically knock out Raptor mice in β cells (βRapKO mice), and mice without RIP-Cre are used as control mice.
[0024] 2. Genotyping
[0025] 1) DNA extraction
[0026] a) Mince the tissue with clean and sterile scissors and add 100ul of AD1 buffer and 25ul of AD2 buffer;
[0027] b) After incubating at room temperature for 10 minutes, place in a metal bath at 95°C for 3 minutes;
[0028] c) Add 100ul of AD3 buffer, mix wel...
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