Does Gene AT5G19490 Play a vital role in seed development? Research Presentation By Ben Torres HC70AL Spring 2004 AT5G19490                             FW >>> RV <<<< 1 1073bp

Did Molecular biology techniques facilitate our quest for knowledge? FW+Jl202 How can we unambiguously tell if there is a T-DNA insert in Gene AT5G19490? RV+Jl202 AT5G19490 Why didn’t the experiment end by picking the super pool with fragments smaller than the gene AT5G19490 on the gel?

What did the AutoRadiograph reveal How was the size of the fragments known? AT5G19490 900bp fragment Did this show unambiguously the smaller fragment was a T-DNA insert?

What do you do When all else fails? cDNA of ortholog gene ATG19490 in the scarlet runner bean SIGnAL "T-DNA Express" Arabidopsis Gene Mapping Tool

Why not do conduct a microarray on the arabidopsis itself? Key for corresponding Stages of plant life On the X axis. 1=wild type ovules 2=wild type 24-hour seeds 3=wild type mid-maturation (Cotyledon stage) 4=wild type mature green seeds 5=wild type post maturation green seeds 6=wild type seedlings (3DAI) 7=wild type leaves 8=wild type roots 9=wild type stem 10=wild type C24 leaves

What about the Salk lines Possible T-DNA homozygous Salk lines Does this show the full genotype? Wild type PCR conducted on salk lines How did the rest of the Genotyping go?

What about the Madison DNA pools PossibleT-DNA insert

What next? Genotype Salk lines using the FW primer ant the LBb1 T-DNA primer correctly to unambiguously define the genotypes Try to obtain a correct sequence from the DNA pool containing the fragment size of interest.