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1. Isolate chromosomal DNA.
2. Digest chromosomal DNA with a restriction enzyme that is unique to the first polycloning site of the P-element vector.
3. Precipitate and re-suspend the ligated DNA (only necessary if transforming by electroporation).
4. Transform bacteria with the ligated DNA. Allow the cells to recover and spread them on plates with appropriate antibiotics. Incubate overnight at 37^oC. Check plates for colonies the next morning.
5. Make an overnight culture of each colony on your plates.
6. Isolate plasmid DNA by an alkaline lysis procedure. Use only 1/2 of your liquid culture for this purpose.
7. Run a test gel of your plasmid DNA. Digest plasmid DNA with the restriction enzyme from the 1st polycloning site and with a restriction enzyme that is unique to the 2nd polycloning site of the P-element vector.
8. Check your digests on an agarose gel. If a valid clone is obtained, freeze the other half of your liquid culture cells.