Transgenic Mouse by Unknown

Author:Unknown
Language: eng
Format: epub
ISBN: 9781493998371
Publisher: Springer US

2 Materials

2.1 MII Oocytes

1.Pregnant mare’s serum gonadotrophin (PMSG).

2.Human chorionic gonadotrophin (hCG).

3.M2 with hyaluronidase.

4.Culture dishes (35 mm, and/or 60 mm) and Nunc 4-well plates.

5.Pipettes and Pipette tips (1000, 200, 10 μl).

6.Operation medium: M2 medium or CZB medium. 20 mM HEPES should be added and NaHCO3 should be decreased to 4.15 mM if M2 medium is made in-house. Cytochalasin B or D (2–5 μg/ml) is added for enucleation.

7.For practical purposes, the most commonly used strains for successful NT cloning for oocyte donors are hybrids, such as B6D2F1 and B6C3F1. The best birth rate was also obtained with the 129 strain, followed by the DBA/2 strain [22]. Females at 6–12 weeks of age give less numbers of oocytes, but they are of a higher quality, compared to the females at 3–4 weeks of age.

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