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Gapdh research august 2009

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Lydia Cortes

The researchers aimed to sequence and analyze glyceraldehyde-3-phosphate dehydrogenase (GAPDH) genes from Cyperus alternifolius and Schefflera actinophylla, two plants endemic to Puerto Rico. They cloned the GAPDH genes from each plant into E. coli bacteria and determined the DNA sequences. Bioinformatics analysis would allow comparison of the GAPDH sequences between the two plants and with other previously sequenced GAPDH genes, helping to understand evolutionary relationships and conserved protein regions. Preliminary results showed successful cloning and transformation of GAPDH genes from each plant species.

Technology
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PCR AMPLIFICATION, CLONING, SEQUENCE DETERMINATION, AND BIOINFORMATICS ANALYSES OF NOVEL PLANT GAPDH GENES FROM CYPERUS ALTERNIFOLIUS , SCHEFFLERA ACTINOPHYLLA AND TROPICAL FLORA ENDEMIC TO PUERTO RICO Lydia E. Cortes and Dr. Michael Rubin Department of Biology and RISE Program University of Puerto Rico at Cayey
GAPDH ,[object Object],[object Object],[object Object],[object Object]
CYPERUS ALTERNIFOLIUS and SCHEFFLERA ACTINOPHYLLA ,[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object]
PROBLEM ,[object Object],[object Object]
SIGNIFICANCE ,[object Object],[object Object]
SPECIFIC AIMS ,[object Object],[object Object],[object Object],[object Object]
HYPOTHESIS ,[object Object]
EXPERIMENTAL DESIGN ,[object Object],1 ul DNA 20 ul of competent cells 42 ° C 90 seconds 2 minutes 950 ul Of SOC media 30 minutes
CONTINUATION OF TRANSFORMATION Amp plates- 900 and 50 ul Overnight (both) LB Broth + Amp solution (100 ul of Amp and 100 ml of LB Broth)
PLASMID PURIFICATION Resuspend in 200ul resuspension solution 250 ul lysis solution 250ul neutralization solution
CONTINUATION PLASMID PURIFICATION Move to tube With column Add 200 ul matrix 2 minutes 500 ul Wash solution 100 ul water 1 minute
SEQUENCE DETERMINATION
CONTINUATION OF SEQUENCE DETERMINATION 10 ul of DNA + 1 ul of primer Pipet 10 ul Seal and mail
BIOINFORMATICS ANALYSIS ,[object Object]
CONTINUATION BIOINFORMATICS ,[object Object],[object Object]
PRELIMINARY RESULTS: CLONING GEL No DNA pGAP Arabidopsis Marker Cyperus Marker Schefflera
PRELIMINARY RESULTS OF TRANSFORMATION Type of DNA Number of cells in 50 microliters Number of cells in 900 microliters CYPERUS ALTERNIFOLIUS 17 440 SCHEFFLERA ACTINOPHYLLA 10 560
ACKNOWLEDGEMENTS ,[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object]
PCR AMPLIFICATION, CLONING, SEQUENCE DETERMINATION, AND BIOINFORMATICS ANALYSES OF NOVEL PLANT GAPDH GENES FROM CYPERUS ALTERNIFOLIUS , SCHEFFLERA ACTINOPHYLLA AND TROPICAL FLORA ENDEMIC TO PUERTO RICO Lydia E. Cortes and Dr. Michael Rubin Department of Biology and RISE Program University of Puerto Rico at Cayey

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Gapdh research august 2009

  • 1. PCR AMPLIFICATION, CLONING, SEQUENCE DETERMINATION, AND BIOINFORMATICS ANALYSES OF NOVEL PLANT GAPDH GENES FROM CYPERUS ALTERNIFOLIUS , SCHEFFLERA ACTINOPHYLLA AND TROPICAL FLORA ENDEMIC TO PUERTO RICO Lydia E. Cortes and Dr. Michael Rubin Department of Biology and RISE Program University of Puerto Rico at Cayey
  • 2.
  • 3.
  • 4.
  • 5.
  • 6.
  • 7.
  • 8.
  • 9. CONTINUATION OF TRANSFORMATION Amp plates- 900 and 50 ul Overnight (both) LB Broth + Amp solution (100 ul of Amp and 100 ml of LB Broth)
  • 10. PLASMID PURIFICATION Resuspend in 200ul resuspension solution 250 ul lysis solution 250ul neutralization solution
  • 11. CONTINUATION PLASMID PURIFICATION Move to tube With column Add 200 ul matrix 2 minutes 500 ul Wash solution 100 ul water 1 minute
  • 13. CONTINUATION OF SEQUENCE DETERMINATION 10 ul of DNA + 1 ul of primer Pipet 10 ul Seal and mail
  • 14.
  • 15.
  • 16. PRELIMINARY RESULTS: CLONING GEL No DNA pGAP Arabidopsis Marker Cyperus Marker Schefflera
  • 17. PRELIMINARY RESULTS OF TRANSFORMATION Type of DNA Number of cells in 50 microliters Number of cells in 900 microliters CYPERUS ALTERNIFOLIUS 17 440 SCHEFFLERA ACTINOPHYLLA 10 560
  • 18.
  • 19. PCR AMPLIFICATION, CLONING, SEQUENCE DETERMINATION, AND BIOINFORMATICS ANALYSES OF NOVEL PLANT GAPDH GENES FROM CYPERUS ALTERNIFOLIUS , SCHEFFLERA ACTINOPHYLLA AND TROPICAL FLORA ENDEMIC TO PUERTO RICO Lydia E. Cortes and Dr. Michael Rubin Department of Biology and RISE Program University of Puerto Rico at Cayey