Generation and isolation of biological materials

1: Seeds for B73, Mo17 and A188 have been planted in the X-8 and X-10 fields.

2: Crosses will need to be performed on two different dates. (Ideally these crosses will utilize plants derived from different plants dates.)

3: Collect ears according the schedule on spreadsheet (appendex 1). From each ear collect three tubeswith eight endosperms, and freeze at -80C. (From 16, 20, and 24 DAP collect four 15 ml tubes full of endosperms.)

4: Do a single DNA prep and a single RNA prep for each ear from the B73, Mo17, B73 X Mo17 and Mo17 X B73.

5: Make cDNA from the RNA and then use 20 IDP primer pairs to screen for allele specific expression for degredation.

The 15ml tubes containingendosperms will be used for large-scale DNA preps and Southern blot analysis. The small preps will be used for PCR markers.