Method for activation of squab, fecundation method for brid egg, and method for hatching out brid embryon containing heteronucleic acid
A technology for heterologous nucleic acid and chick, applied in the field of bird egg activation, which can solve cumbersome problems and other problems
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Embodiment 1
[0121] 1. Freshly laid eggs of 43 unfertilized Barred Rock chickens were disinfected by wiping the shells with 3% hydrogen peroxide, and then placed on racks.
[0122] 2. Within 1 hour before insemination, on the same morning, obtain semen from 4 isolated Rock roosters and pack into in vial.
[0123] 3. Concentrate the semen of 4 roosters and mix with 1ml mix.
[0124] 4. Use No. 1 and No. 22 needles to mix the semen and The mixture was drawn into a 1ml syringe.
[0125] 5. Use a needle to make an opening at the large, blunt end of the egg shell, and pass the needle through the opening at an angle of 15 to the surface of the egg shell.
[0126] 6. Pass the needle through the air cell until its tip just penetrates the membrane surrounding the yolk and blastodisc.
[0127] 7. Inject a 0.05 ml drop of the sperm sample onto the surface of the yolk adjacent to the membrane.
[0128] 8. Remove the needle attached to the syringe from the egg.
[0129] 9. Use a small piece o...
Embodiment 2
[0134] The following data are from commercial "Hy-Line Variety Brown" hens that lay brown eggs:
[0135] 1. Collect 270 newly spawned eggs at 6:30 AM.
[0136] 2. The semen collected from the male Black Giant was immediately diluted 50:50 with the diluent.
[0137] 3. Within 20 minutes after sperm collection, send the sperm and eggs to the laboratory for experiments.
[0138] 4. The eggs were divided into two groups, the experimental group and the negative control group, 135 eggs in each group.
[0139] 5. Inject the experimental eggs with 10 μl of diluted semen as described previously.
[0140] 6. The negative control group was not injected.
[0141] 7. As previously described, the injected eggs are sealed with a silicone sealant and placed in an incubator to hatch.
[0142] After 8.5 days, check the fertility and make a record.
[0143] 9. Of the 135 eggs in the experimental group, 35 (24%) were fertile. None of the eggs in the control group showed signs of development...
Embodiment 3
[0144] Example 3: Parthenogenesis
[0145] 1. Collect 270 newly spawned eggs in the morning.
[0146] 2. Eggs were divided into two groups, the experimental group and the negative control group, with 135 eggs in each group.
[0147] 3. Make a 2-3 mm hole through the shell and shell membrane so that the yolk can be seen.
[0148] 4. Gently "prick" and / or penetrate the membrane surrounding the yolk and blastodisc, taking care not to break the yolk. (Alternatively, openings in the egg shell and penetration of the vitelline membrane can be performed simultaneously by inserting a 25 gauge syringe needle through the egg shell and into the yolk).
[0149] 5. Seal the treated eggs with a sealant and put them into an incubator to hatch.
[0150] 6. Check the development of eggs on day 5 and record the development ratio before returning to the incubator.
[0151] 7. Allow the eggs to continue to develop until the chicks hatch, taking note of hatch percentage and health conditions. ...
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Abstract
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