Embryo Cryopreservation or Rederivation via Natural Matings using PI’s Males and Females
Transfer mice
to TMF
PPMS
Week 1
2-cell
collection
Week 2
Quality
control test
Week 3
Genotype
Pups
Week 6-7
Quick Guide (see below for more detailed flow chart):
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PI submits a project request form in PPMS
PI transfers 10 or more 10-20 week old males and 3 week old females to Peds G08 (week1)
Females are superovulated and mated, and 2-cell embryos are collected (week 2)
For Rederivation – 2-cell embryos are implanted into recipient females (week 2)
For Cryopreservation – 2-cell embryos are cryopreserved (week 2)
TMF thaws one straw for quality control and implants embryos into recipient female (week 3)
PI genotypes toe biopsies from pups born (from both cryo QC thaw and rederivation) and sends results to the Core (week 6-7)
For Rederivation - Mice are weaned onto PI’s smart key and protocol number and transferred to PI’s housing location.
For Cryopreservation - Mice are euthanized or weaned onto PI’s smart key and protocol number and transferred to PI’s housing
location.
Detailed Guide:
Before Project Begins:
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PI is responsible to order the project from PPMS
PI needs contact TMF to discuss project
PI is responsible to transfer mice to Peds G08
Project Flow:
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PI lab is responsible for transferring mice of more than 10 pairs to Peds G08.
For male mice: proven breeder of approximately 10 to 20 weeks.
For female mice: young female mice of approximately 3 to 6 weeks of age.
TMF will superovulate, mate, and collect 2-cell embryos from the female mice
For Rederivation: TMF will transfer 2-cell embryos into recipient females and send the PI an update.
For Cryopreservation: TMF will cryopreserve the 2-cell embryos. One straw strain will be thawed for a quality control test and the
thawed embryos will be implanted into a recipient female
Toe clips will be taken from the pups for genotyping when they are 7 days old
PI lab is responsible for screening pups from toe biopsies provided from TMF
PI lab is responsible to send TMF screening result and copy image for documentation
When pups are 4 weeks old, TMF will use PI’s IACUC protocol and smart key to wean the pups
For rederivation: TMF will send recipient moms for Pathogen testing through DAR, PI is responsible to pay testing fee.
For Cryopreservation: TMF will Euthanasia pups or transfer to PI if requested
For Rederivation: When pathogen results come back negative, TMF will arrange transfer of rederived mice to PI lab, PI lab is
responsible for checking that the mice have been received.
Service Rate Structure:
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TMF will bill PI on the month of the project initiated:
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Rederivation fresh embryo fee: $1067
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Embryo Cryopreservation fee: $1705
Extra fee will be billed to PI lab in the following cases:
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$173.67 for every two recipient moms for pathogen test
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Per diem fee after pups wean
$38 per hour plus material cost when TMF extract DNA from tail samples
Mouse shipping costs if rederived pups are being transferred outside of Emory
Project Guarantee and Quality Control
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TMF will guarantee the existing Core protocol and Core SOP are followed, and will follow any protocols provided from PI