DNA Pronuclear Microinjection
Investigators who have transgene constructs ready should complete a DNA Pronuclear Microinjection Request Form. The microinjection will be scheduled after we have received the construct DNA and the completed request form.
The full service includes DNA purification, microinjection and housing potential transgenic pups until three weeks old. After this point, the investigator is responsible for colony management and housing charges. We guarantee three transgenic mice or 50 pups whichever comes first.
At least 60 ug of circular construct DNA is needed. Please use Endofree plasmid kit, clontech nucleobond midiprep or CsCl-Ethidium Bromide gradients method to prepare your plasmid DNA. For BAC DNA please follow the BAC preparation protocol.
DNA is to be digested by the Investigator prior to delivery to the CMGM. Please run a sample on a gel and supply the picture with the DNA.
You can ship your DNA sample by FedEx at ambient temperature by adding 1/10 volume of NaAc 3M pH5.2 and 2 volume of EtOH 100% into your sample. Please seal the 1.5 ml tube well, put it into a 15 or 50 ml tube and then put it in an air bubble envelope. A small tube could be crushed easily during the shipment.
Timeline for DNA pronuclear microinjection:
|Request Submitted||Microinjection||Pups born||Pups weaned, tails cut|
|3 to 4 weeks - order mice, superovulation, DNA purification||3 weeks - pregnancy||3 weeks|