8) Micro Bio Lab Practical (HurBi biplate & Genetics: Transformation)

What is the purpose of the HurBi biplate urinalysis?

To grow bacteria from a urine sample, Determine whether bacteriuria are present, Presumptively identify the cause

What % of UTI’s are caused by E. coli?

90%

What is chromagar?

Agar that turns different colors based on differences in species-specific metabolism

What are the rules regarding the colony count in HurBi biplate?

less than 10 colonies = less than 10,000 bacteria = negative bacteriuria

10-100 colonies = 10,000-100,000 bacteria = borderline

More than 100 colonies = more than 100,000 bacteria = positive bacteriuria

How to calculate the Original Cell Density (OCD)?

OCD = Colony forming units (CFU) / loop volume = CFU x 1000

What is the purpose of the genetics transformation?

To transform ampicillin sensitive E. coli DH5-alpha into an ampicillin resistant strain through plasmid uptake

What is DNA transformation?

The process where an organism takes up foreign DNA from its surroundings

What is a plasmid?

Small, circular DNA that is not part of an organism’s chromosome

What do you need to do to calcium chloride in transformation experiment?

Incubate at ice cold temp.

What needs to be done to the cell w/ plasmid DNA during transformation test?

Heat shock at 42C

What needs to be done with E. coli in transformation test?

Spread on plates w/ selective growth media