How to Make a Transgenic Mouse with Emory’s Transgenic Mouse Core Facility:
PPMS
Deliver
Vector
Injection
Genotype
Toe
Confirm +
Genotype
with ear
Week 1
Week 3
Week 7
Week 8
Transfer
Founders
Week 11
Quick Guide (see below for more detailed flow chart):
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7.
PI creates an account in PPMS (if doesn’t already have one) and submits a project request form.
PI meets with Core Director to discuss project and deliver your vector to the Core
The Mouse Core purifies the transgene and orders embryo donor females (weeks 1-2)
Construct is injected by the Core (week 3), pups are born 3 weeks later
PI genotypes toe biopsies from pups born and sends results to the Core (week 7).
PI genotypes ear biopsies from pups identified as positive to confirm TG+ genotype
Mice are weaned onto PI’s smart key and protocol number and transferred to PI’s housing location (week 11).
Detailed Guide:
Before Project Begins:
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PI needs to meet with the Core staff for a free consultation to go over the transgenic construct design and screening strategy
PI is responsible for building the transgenic construct and developing the screening strategy, showing transgene amplification from
one copy number
PI is responsible to submit the following to TMF:

Vector map and screening strategy in electronic version

Gel image showing transgene detection for one copy number

30 ug uncut plasmid DNA, or at least 50 ul of minimum 5 ng/ul of purified linearized BAC DNA
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Enzyme(s) to release the transgene from plasmid DNA
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Gel image showing clear transgene release with designated enzyme(s)
PI is responsible for entering a project request form into PPMS. A valid smart key and protocol number must be provided.
External PI’s must submit a Purchase order for the price of the service they are requesting before the project can begin
Project Flow:
1.
For plasmid DNA, TMF will cut the vector with enzyme to release transgene and purify the vector, check the concentration, and make
dilutions for injection.
2. TMF will inject the construct and send the PI an update.
3. TMF will notify PI lab when pups are born.
4. TMF will collect toe biopsies at 7 days of age from pups born and give to PI. If PI is off campus, we will ship biopsies on ice using
PI’s FedEx account number.
5. PI is responsible for screening the samples and sending the results to TMF.
6. Once results are received by TMF and the mice are at least 14 days of age, we will collect ear biopsies from potential transgenic
founders for confirmation and give to the PI lab.
7. PI lab is responsible for screening the ear biopsies to confirm the genotype and sending the results to TMF
8. At 4 weeks of age, TMF will wean transgenic founders onto PI’s smart key. TMF will sack negative mice when the positives have
been confirmed
9. TMF will arrange the transport of the transgenic founders to the PI’s housing location, notifying the lab of the transport.
10. PI lab is responsible for verifying that the lab has received the mice
Service Rate Structure:
1.
TMF will bill PI on the month of the project initiated:

$4320 for Transgenic Mouse Production: B6
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$4120 for Transgenic Mouse Production: DB6F2 Hybrids
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$4120 for Transgenic Mouse Production: FVB
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$5187 for BAC TG Mouse Production: FVB
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$5805 for BAC TG Mouse Production: C57Bl/6
2.
Extra fee will be billed to PI lab in the following cases:
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If TMF is requested to purify DNA from tail or ear biopsies, $4 per tail
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If shipping charges are incurred transporting mice to external PIs
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If per diem fees are incurred after weaning and awaiting transport, for external PIs
Project Guarantee and Quality Control
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TMF will guarantee at least 2 transgenic founders
TMF will not guarantee transgene expression
TMF will guarantee the existing Core protocol and Core SOP are followed