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BISC110: Series 4 Lab 11 Enzymology: Difference between revisions
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10. You are now ready to begin your assays. Take your rack of 8 tubes, a 1mL pipette, your timer, your 3mM sucrose solution and your notebook to one of the 55°C water baths. Put your tubes into the water bath. Then, every 30sec, add 0.5mL of the 3.0mM sucrose solution to the next tube and return that tube to the water bath. Follow the timing listed in the table below. | 10. You are now ready to begin your assays. Take your rack of 8 tubes, a 1mL pipette, your timer, your 3mM sucrose solution and your notebook to one of the 55°C water baths. Put your tubes into the water bath. Then, every 30sec, add 0.5mL of the 3.0mM sucrose solution to the next tube and return that tube to the water bath. Follow the timing listed in the table below. | ||
{| border="1" | |||
|+ | |||
! Tube# !! Contents !! Add 0.5ml <br>of 3mM Sucrose !! Add 3ml <br>of DNS reagent | |||
|- | |||
| 1 | |||
| water | |||
| do not add | |||
| 14:00 min. | |||
|- | |||
| 2 | |||
| glucose | |||
| do not add | |||
| 14:30 min. | |||
|- | |||
| 3 | |||
| sucrose | |||
| 00:00 | |||
| 15:00 | |||
|- | |||
| 4 | |||
| sucrose + invertase | |||
| 00:30min. | |||
| 15:30min. | |||
|- | |||
| 5 | |||
| WT yst/2.0% glucose | |||
| 01:00 | |||
| 16:00 | |||
|- | |||
| 6 | |||
| WT yst/0.05% glucose | |||
| 01:30 | |||
| 16:30 | |||
|- | |||
| 7 | |||
| MUT yst/2.9% glucose | |||
| 02:00 | |||
| 17:00 | |||
|- | |||
| 8 | |||
| MUT yst/0.05% glucose | |||
| 02:30 | |||
| 17:30 | |||
|- | |||
|} | |||
11. After each addition, you should vortex the tube briefly to mix the contents, then move the tube to another place in your rack in the 55°C bath to avoid accidentally adding sucrose twice to the same tube. After your sucrose additions are completed, please move away from the water bath to give other groups room to work. | 11. After each addition, you should vortex the tube briefly to mix the contents, then move the tube to another place in your rack in the 55°C bath to avoid accidentally adding sucrose twice to the same tube. After your sucrose additions are completed, please move away from the water bath to give other groups room to work. | ||
