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Team:BYU Provo/Notebook/SmallPhage/Winterexp/Period3/Exp/4.3 Top Agar Stock Preparation
From 2013.igem.org
| Small Phage March - April Notebook: Experiments
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4.3 Top Agar Stock Preparation
I) Purpose - Learn the procedure for making top agar - Producing top agar stock for use II) Expected outcome - Produce stock top agar (at ×2 concentration) for phage selection.
III) Proposed procedures 1. LB broth: Make 250mL of LB broth following the instruction provided by the manufacturer. 2. 3.2% stock: Dissolve 5g LB broth powder and 8g agar powder in 250mL of distilled water. 3. 2.4% stock: 90mL of the 3.2% stock + 30mL of LB broth 4. 1.6% stock: 100mL of the 3.2% stock + 100mL of LB broth 5. 0.8% stock: 100mL of the 1.6% stock + 100mL of LB broth (normal ×2 stock top agar) 6. Stock solution should be put in glass jars and autoclaved 30 minutes, liquid cycle 7. When ready for use, warm stock agar till it’s liquid and then do a 1:1 dilution with LB broth to produce the top agar at the concentration ready for use IV) Actual procedures - (April 3) Made concentrated top agar stock and LB solution, autoclaved and should be ready for dilution/use
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