undergrads 2

We are performing PCR to identify if 35S:NF-YB12:YFP:HA was ever inserter into the genome. We will be amplifying the region using a 35S forward primer and a gene specific reverse primer. This should tell us if the transgene made it into the genome. We could also amplify another region, using a YFP reverse and gene specific forward primer combination. We could amplify another region, using the primer combination of gene specific forward and HA reverse. One final combination, which would give you the answer, but would be a sloppy approach is using the two primer extremities of the construct, 35S forward and HA reverse. This requires doing a little math for confirmation. This requires looking at the sequence of the construct, counting nucleotides for each portion of the construct, and adding this together to get a collective base count of the construct. The amplified band should be exactly that size.