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Superovulation Protocol | Taconic Biosciences

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Superovulation is a technique used to get female mice to release more eggs than normal. It is a critical step in collection of morulae and/or blastocysts which can be used as hosts for genetically modified embryonic stem cells (ESCs) during generation of genetically modified mouse models. Morulae are collected at 2.5 days post coitus (dpc) and incubated in vitro overnight for use in injections the next day. Alternatively, blastocysts are collected at 3.5 dpc and used for injections shortly after collection. The injected blasts are implanted in pseudopregnant foster mothers and chimeric pups consisting of genetic contributions from both host blastocyst as well as the embryonic stem cells are born. C57BL/6 mice (B6) are commonly used as blastocyst donors for ESC injection. Below is a protocol developed by Taconic to work well with strains on the Taconic C57BL/6NTac background. For use with the following Taconic models: Hormone reconstitution: Light cycle: 12hrs light / 12 hrs dark Animal

Hormone reconstitution: PMSG is reconstituted from a lyophilized powder at an activity of 2000 IU per vial by adding 2 ml of sterile PBS or sterile 0.9% saline. 50 microliters are then aliquoted into 2 ml conical tubes (totaling approximately 40 tubes per vial). Tubes are labeled and placed in 80 °C freezer. HCG is reconstituted from a lyophilized powder at an activity of 10,000 IU per vial by adding 10 ml of sterile PBS or sterile 0.9% saline. 50 microliters are then aliquoted into 2 ml conical tubes (totaling approximately 200 tubes per vial). Tubes are labeled and placed in -80 °C…

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