Purpose:
To show how genes can be regulated by using the lac operon as an example.
Materials:
BL21 (DE3) + egfp (un-induced)
3 agar plates with Kanamycin
0.5 mM IPTG
Methods:
Plate 2 kanamycin plates with un-induced BL21(DE3) + egfp.
Label plates no IPTG and IPTG
Place in 37°C incubator overnight
To turn gene on for expression
Plate 1: Do not add anything
Plate 2: Using a sterile pipette tip, add 8 µL 0.5 mM IPTG per colony.
Allow IPTG to soak in
Incubate at 37°C overnight
Place the plate under UV light to see bacteria glow.
Calculation: To make 0.5 mM IPTG, mix 50 µL 1.0 M IPTG + 50 µL LB broth
This would be a cool extension of this lab that might be more relatable to the students.
Lab by Ingrid Coulson,
9th grade Honors Biology teacher
Broughton High School, Raleigh, NC
To show how genes can be regulated by using the lac operon as an example.
Materials:
BL21 (DE3) + egfp (un-induced)
3 agar plates with Kanamycin
0.5 mM IPTG
Methods:
Plate 2 kanamycin plates with un-induced BL21(DE3) + egfp.
Label plates no IPTG and IPTG
Place in 37°C incubator overnight
To turn gene on for expression
Plate 1: Do not add anything
Plate 2: Using a sterile pipette tip, add 8 µL 0.5 mM IPTG per colony.
Allow IPTG to soak in
Incubate at 37°C overnight
Place the plate under UV light to see bacteria glow.
Calculation: To make 0.5 mM IPTG, mix 50 µL 1.0 M IPTG + 50 µL LB broth
This would be a cool extension of this lab that might be more relatable to the students.
Lab by Ingrid Coulson,
9th grade Honors Biology teacher
Broughton High School, Raleigh, NC
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