VPB 321: Animal Biotechnology (2+1)

Steps in prodction of transgenic animals by DNA micro injection

STEPS IN PRODUCTION OF TRANSGENIC ANIMALS BY DNA MICRO INJECTION 

• 4- 6 week old female mice are superovulated by administering 5 IU each of PMSG, HCG and mated to male mice.
• Pronuclear stage – eggs are recovered after 12 hrs.
• Pronuclear transgene – DNA construct is injected into the male pronuclei by using micromanipulator.
• Embryos after microinjection are transferred to recipients and allowed to develop to term.
• Integration of the foreign gene in the new born animals could be confirmed by techniques like
• Polymerase chain reaction
• Southern hybridization
• In situ Hybridization.