Table IX. Summary of the volume of chemicals to be added in different tubes.
Tube
DNA Buffer EcoRI BamHI Hind III Water
(clone)
E
8 µL
1 µL
1 µL
-
-
-
B
8 µL
1 µL
-
1 µL
-
-
H
8 µL
1 µL
-
-
1 µL
-
C
8 µL
1 µL
-
-
-
1 µL
*All groups share the same BamHI, EcoRI, HindIII enzymes, buffer, and water.
3. Pool reagents and mixed by tapping the tube bottom on lab bench, or with a short pu
in a microcentrifuge.
4. Incubate all reaction tubes for a minimum of 60 minutes at 37°C. You may need to
incubate the reactions for a longer period (you're your instructor asks for).
B: Cast Agarose Gel
1. Seal ends of gel-casting tray with tape, and insert well forming comb. Place gel-
casting tray out of the way on lab bench, so that agarose poured in next step can set
undisturbed.
2. Dissolve 1 g agarose per 100 mL of TBE buffer by heating in a microwave oven. C
agarose solution (1%) on to about 60°C.
