1. At5g65640 - A mutant phenotype? Emily Eder HC70AL - Spring 2005

2. Arabidopsis Gene At5g65640 Plant genotyping Sequencing and T-DNA insert First observation of mutant phenotypes Nomarski RNA extraction – microarray – expression

3. Genotyping 4 Homozygous, 0 Heterozygous, and 8 Wild Type Plants Homozygous

4. Sequencing the Mutant Band Why is it necessary to sequence the band? We must verify the T-DNA insertion site

5. AT5g65640 2026 Base Pairs in Length Codes for basic helix-loop-helix (bHLH) protein 146

6. Mutant Phenotypes

7. Mutant Phenotypes cont.

8. Using Microscope to Examine Phenotypes

9. Microscope Examination cont.

13. Mutant Nomarski

14. Wild Type Nomarski

15. Using Gel Electrophoresis to Determine RNA Expression

16. Gel Electrophoresis to Determine RNA Expression

17. Using a Genechip (Microarray) to Determine RNA Expression

18. Arabidopsis Gene AT5g65640 4 Homozygous Mutant Plants - all with visible phenotype 1 T-DNA insert - forward orientation RNA expression revealed a large amount of RNA present in the seed

19. What’s next? Can we be sure that this is an actual mutant phenotype? More research is necessary