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

2. 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?

3. 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?

4. 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

5. 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

6. 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?

7. What about the Madison DNA pools
PossibleT-DNA insert

8. 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.