By: Erick G. Rodríguez & Jefry Lopez

The main purpose of this experiment is to isolate and

sequence the GAPDH (glyceraldehyde-3-phosphate

dehydrogenase) gene from the Cucumis sativus plant.

GAPDH

An enzyme

Catalyzes the sixth step of glycolysis

Serves to break down glucose for energy and carbon molecules.

Most abundant enzyme in cells

Dicot

Grows up by wrapping

around objects using

spiraling tendrils.

Its fruits are called

cucumbers.

Kingdom: Plantae

Division: Magnoliophyta

Class: Magnoliopsida

Order: Cucurbitales

Family: Cucurbitaceae

Genus: Cucumis

Species: C. sativus

Nucleic Acid Extraction Separate DNA from other “waste components” of the cell.

GAPDH PCR

Amplify a specific segment of the GAPC gene by utilizing repeated

cycles of DNA synthesis.

Electrophoresis Separate DNA fragments by size using an electric current that

passes through the agarose gel.

PCR Purification

To clean DNA from all the impurities that result from the PCR

using Chromatography.

Ligation

To join two piece of linear DNA to form one single piece.

Catalyzed by an enzyme called DNA ligase.

Transformation

Used to introduce the plasmid into living bacterial cells so it can be

replicated

Plasmid purification

Perform a miniprep of the plasmid DNA in preparation

for DNA sequencing.

• DNA SequencingDetermine the exact order of nucleotide bases.

• Compare our results with the other data bases

Plasmid purification

Perform a miniprep of the plasmid DNA in preparation for

DNA sequencing.

•DNA SequencingDetermine the exact order of nucleotide bases.

•Bioinformatics

Compare our results with the other data bases

Future steps

Conclusion

This experiment was not completed because the bacteria

needed for the DNA purification didn’t grew. One of the possible

explanations for it is that because of the Amphiciline this

bacteria didn’t grew. So the next steps couldn’t be completed.