LAB FINAL

What type of chromatography separates molecules based on their sizes?

Gel filtration

What type of chromatography separates molecules based on their charges?

Ion exchange chromatography.

What type of chromatography separates molecules based on hydrophobic interaction?

Reverse-phase chromatography

What type of chromatography has the highest specificity?

Affinity chromatography,

What reagents are required for PCR?

• PV92 primer mix

• Cheek cell DNA sample (template DNA)

• PCR reaction pellet

• Nuclease-free water

What type of DNA sequence in the human genome is particularly useful for DNA fingerprinting?

Alu elements (SINEs, Short Interspersed Elements)

What is bacterial transformation?

The process by which bacteria take up foreign plasmid DNA. In this lab, E. coli is transformed with the pGREEN plasmid

What type of DNA is used in transforming bacteria?

Plasmid DNA (pGREEN plasmid)

What is a selectable marker? Why was penicillin added to the plate?

A selectable marker is a gene on the plasmid that allows transformed bacteria to survive under selective conditions and the LB/Amp (ampicillin) plates select for bacteria that have taken up the plasmid because only bacteria carrying the antibiotic-resistance gene will grow.

What is the temperature used for heat shock of bacteria? How long do you need to heat shock bacteria to get the best result?

42°C for 90 seconds.

How long do you need to culture bacteria before you can see colonies on the plate?

24–36 hours at 37°C (or 48–72 hours at room temperature)