Related link about transgenic mice in Creative Animodel： http://www.creative-animodel.com/Animal-Model-Development/Transgenic-Animals.html

1. Related link about transgenic mice in Creative Animodel：
http://www.creative-animodel.com/Animal-Model-Development/Transgenic-Animals.html
Preparation of Transgenic Mice
Tags: transgenic mice, injection, receptor, donor
1. To get a 7~8 weeks old female mouse as the donor, closing the vaginal mouth. At 3:00 pm,
giving PMSG (10 IU) intraperitoneal injection to each mouse.
2. 47~48 hours later, each mouse is given intraperitoneal injection of hCG (0.8 IU) and combined
with normal male rats in one cage. Getting a number of female mice (2 months or above) as
receptors, letting them stay with the ligation male rats in one cage.
3. The second day before 9:00, observing the donors, receptors, keeping sperm suppository for
spare. Picking receptor cages out with isolation measures.
4. Around 10:30, the donor should be killed by breaking the neck and the whole oviduct is
removed to hyaluronic acid~0.3mg/M2 liquid. Under the microscope, tearing the ampulla of
fallopian tube with tweezers. Fertilized eggs outflow together with the particles cells.
5. Carefully observing the fertilized eggs in the hyaluronidase M2 liquid. When the granulosa
cells surrounded fertilized eggs are falling off, we should suck them out and wash them in M2
liquid, and finally placed them in M16 medium with 5% CO2 to culture under the environment of
37C0.
6. Under the microscope, selecting the fertilized eggs with full cells, transparent strap and visible
male pronucleus.
7. Installing egg needle and injection needle, making the end parallel to loading platform.
Dropping one drop of M2 in the central of concave slide, covering paraffin oil and moving
fertilized eggs in. The DNA bubbles in the injection pin should be all previously eliminated.
8. Under microscope of high magnification, making the injection needle touch the fallopian tube,
letting DNA flow out slowly and controlling the air flow. Repeatedly sucking and blowing the
fertilized egg, making it stay in the best position. Injecting needle into the male pronucleus of
fertilized eggs until seeing its enlargement. Placing the injected and the non-injected eggs
separated, avoiding mixing. After injection, adding 5% CO2 and keeping the fertilized eggs in
incubator with 37C0 statement.
9. Giving an Anesthesia to the donor, injecting 1% sodium 0.01ml/g to the intraperitoneal part.
Connecting fallopian tube and fixing with fat tweezers. Finding the fallopian tube opening under
the microscope. Getting the survival fertilized eggs after culture. The method to get is to suck a
longer period of M2, after a bubble absorption and then absorbing fertilized eggs. Trying to have
close space, a section of liquid, a bubble absorption and another section of liquid. Totally, four
sections of liquid and three bubbles. Except for the longer period of liquid, the remaining liquid is

2. about 1cm long, and the bubble is about 0.2cm long. Inserting the transplanted nozzle into the
fallopian tube, gently blowinng the liquid into the tube graft. When seeing fallopian tube pot
abdominal enlargement and three bubbles clearly, it means a successful transplantation. Returning
the ovaries and fallopian tubes to the abdominal cavity, suturing the muscles and skin.
10. The receptor should be weighed once a week. When the weight increases, you can determine
an initial pregnancy. 19~21 days after the operation, the rats start to delivery. 3 weeks after that,
shearing ear, listing serial numbers, shearing tail and detecting.
The original article: http://www.qmed.com/news/supplier/preparation-transgenic-mice