Transgenics DNA Purification for Microinjection Service

If the client chooses to have the VARI core facility purify the DNA construct, then the following procedure should be followed:

1. The gene of interest should be cloned into a plasmid vector and the plasmid purified by CsCl gradient or using the Qiagen plasmid purification system.
2. Excise the DNA fragment of interest from the plasmid vector by restriction enzyme digestion. It is necessary to provide VARI with 10 - 15 µg of the digest.
3. Analyze an; aliquot of this restriction digest by gel electrophoresis and photograph the gel. This gel must contain DNA size markers for reference.
4. On the photograph, mark the size of the markers and mark the band containing the DNA fragment to be isolated for microinjection.
5. Ship the restriction enzyme digest on dry ice to the VARI transgenic lab.