18.3: Pull Sheets/Materials
- Page ID
- 118041
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LIVE MATERIAL
Escherichia coli - MM294 - 2 slants (or plates)
- Carolina #21-1530
- Give to Micro tech when they arrive. They will need to make a total of 12 E. coli streak plates for this lab (we need 6 streak plates per section)
- Note: this is not the same E. coli we use in Biol74 – this MM294 strain takes up plasmids more readily
Plasmid pAmp – need 2
- Carolina #21-1430
- (if we have enough $, can we order 1 extra to account for student pipetting errors? – if not, that is okay – cost is $188/tube)
EQUIPMENT
| Item | How many? |
| Gloves (s, m, l) | 1 box of each |
| Goggles | 30 |
| Inoculating loops | 6 (prefer 12 if available) |
| Spreading rods | 6 (prefer 12 if available) |
| Bunsen burners | 6 |
| Strikers for Bunsen burners | 6 |
| Eppendorf tube racks | 6 |
| Plastic beakers for ethanol | 6 |
|
Ice buckets OR Dyna Chill ™ lab coolers (frozen before lab) |
7 (6 for students, 1 for instructor plasmids) |
|
Hot water bath – set to 42°C with trays in water bath to hold microcentrifuge tubes |
1 |
| P20 micropipettes (0-20ul) | 6 |
| P100 micropipette s (measure to100ul) | 6 |
| P200 micropipettes (measure to 200ul | 6 |
| Sterile micropipette tips (fit to micropipettes above) | 6 boxes of each type |
| Sterile microcentrifuge tubes | 40 |
|
Large waste beakers or jars Each labeled “Used Eppendorf Tubes & Pipette Tips” |
3 (one per bench) |
CHEMICALS & SOLUTIONS
| Item | How many? |
|
2.0 ml 0.05 M CaCl2 – in 250 µl Eppendorf tubes At least 250uL 0.05 M CaCl2 in each Eppendorf tube x 28 Eppendorf tubes (14 per section) |
8 - keep cold & sterile |
| 95% Ethanol - keep cold | 200 ml in 2 Erlenmeyer flasks |
|
2.0 ml Luria broth – in 250 µl Eppendorf tubes At least 250uL Luria broth in each Eppendorf tube x 28 Eppendorf tubes (14 per section) |
8 - keep cold & sterile |
|
LB/Amp agar plates Carolina #21-6621 (ready to pour, give to micro tech) |
need 28 plates across 2 lab sections |
|
LB agar plates (micro tech may be able to make) |
need 28 plates across 2 lab sections |
| Spray cleaner or wavicide | 3 bottles |
| Paper towels | 3 rolls (or use those available in the lab room) |
Note: After lab: Incubate inoculated Petri dishes at 37° C for 24 hours; then move to 5° C cooler.
DAY 2
| Item | How many? |
| Spray cleaner or wavicide | 3 bottles |
| ‘Scotch’ (clear) tape or glue sticks (make sure they are fresh) | 15 |