GFP Purification Process Assessment

This video tutorial from Edvotek covers the purification of Green Fluorescent Protein (GFP) using column chromatography. It details the process of packing and equilibrating a chromatography column with a molecular sieve matrix, running the GFP-containing lysate through the column, and collecting the fluorescent fractions. The video also explains how to monitor the progress of GFP using UV light and how to prepare samples for further analysis in Module IV.

What is the primary purpose of Module III in the GFP purification process?

• To synthesize GFP in the laboratory

• To purify GFP using column chromatography

• To analyze GFP using SDS Gel Electrophoresis

• To degrade GFP for further study

Which of the following is NOT required for the GFP purification process in Module III?

• DNA polymerase

• Screw top microcentrifuge tubes

• Microtiter plate

• UV light source

What is the first step in packing and equilibrating the column?

• Mix the Molecular Sieve Matrix

• Add elution buffer to the column

• Place an empty beaker underneath the column

• Vertically mount the column onto a ring stand

How much of the mixed slurry should be pipetted into the column?

• 2 ml

• 4 ml

• 3 ml

• 1 ml

What should be done if the packed matrix is too low in the column?

• Increase the column temperature

• Remove some of the matrix

• Add more elution buffer

• Add additional slurry

What is the initial volume of GFP extract loaded into the column?

• 0.1 ml

• 0.2 ml

• 0.3 ml

• 0.4 ml

How should the progress of GFP in the gel matrix be monitored?

• By illuminating with a long-wave UV light source

• By using a thermometer

• By checking the temperature

• By measuring the pH

How many drops per well should be collected in the microtiter plate?

• 2 drops

• 3 drops

• 4 drops

• 5 drops

What should be done with the brightest elution identified in the microtiter plate?

• Discard it

• Heat it to 100°C

• Transfer it to a screw top microcentrifuge tube

• Mix it with another sample

What is the next step after completing Module III?

• Begin Module II

• Repeat Module III

• Proceed to Module IV

• Start Module I