a timed presentation - do not click the mouse. approximate run time - 4 minutes
TRANSCRIPT
A Timed Presentation - do not click the mouse.
Approximate Run Time - 4 minutes
Gel is 22.5 x 22.5 cm in size
Comb produces 28 wells
Uncut: DNA 18.75 μl10 x Buffer 15 μlWater 116.25 μl 6 x Loading Buffer 30 μl
30 μl/well – approximately 1.25 μg DNA/well
Single cut: DNA 37.5 μl10 x Buffer 22.5 μl
BamHI 15 μlWater 150 μl 6 x Loading Buffer 45 μl
30 μl/well – approximately 1.67 μg DNA/well
Bouble cut:DNA 37.5 μl10 x Buffer 22.5 μlBamHI 11.25 μlEcoRI 11.25 μ lWater 142.5 μl 6 x Loading Buffer 45 μl
30 μl/well – approximately 1.67 μg DNA/well
Each sample of DNA was incubated for 1 hour at 37°C.
Each sample and the ladder was then loaded into each well as outlined in the following slides.
pBabe/cpp326000 bp
BamHI
EcoRI
900 bp
5100 bp
pBabe plasmid donatedby Dr. Tang
Lane 1
Lane 10
Lane 20
Lane 28
2
3
1
BufferStain
27
17
LadderRows:
1
The SBI4U class ofHill Park Secondary School
wish to thank
Dr. D. Tang
(McMaster University/St. Joseph’s Hospital)and his lab technician
for donating the DNA, Restriction Enzymesand Ladder and for calculating the amounts
of material required to complete this procedure successfully.