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Transgene Microinjection 

  • Standard plasmid transgene in F1-hybrid strain: $ 2100
  • BAC transgene in F1-hybrid strain: $ 2300
  • Inquire about special requests: transgene injections in inbred B6 strain.

For all services, download the Service Order Form and submit to the GMF Administration via email: hdemanbey@mcb.harvard.edu or to Biolabs room 1062. Arrange with OAR for transfer of animal to GMF; form can be downloaded here. Once submitted, purify transgene DNA as per instructions on the protocol and then submit transgene DNA with the sample sheet and signed Order Form to the Administrative Office.

GMF Responsibilities:
  • Inject 200 - 300 embryos, produce > 30 offspring animals from injected embryos.
  • Wean offspring, tag and tail, ship tail samples to investigator for genotyping.
  • Transfer transgenic founder animals to PI colony.

 

Transgene Microinjection Tasks and Approximate Timeline

Tasks Timeline Responsible Parties

 

Transgene Construction





Variable

6 - 8 weeks



1) Design and clone transgene vector.
2) Purify transgene insert.
3) Schedule transgene injections with GMF.

 

Pronuclear Microinjection



Start t = 0

6 - 9 weeks



1) Microinject around 300 zygotes
2) Produce minimal number of 30 offspring
3) Wean offspring, transfer to investigator

 

Genotyping Offspring



Start t = 9 weeks

18 - 36 weeks


1) Genotype founder animals
2) Expand, breed transgenic colony
3) Produce animals for analysis

 

 

Analysis of Transgenic Mice

G0 : Analysis as early as a few days after injection.

F1 - Fn : Analysis up to several years


Analysis of transgene expressing mice

1) G0 - mice during gestation or postnatally
2) Expression and phenotype analysis in F1 to Fn mice