Team:BYU Provo/Notebook/SmallPhage/Winterexp/Period3/Exp/4.3 Top Agar Stock Preparation

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'''IV) Actual procedures'''
'''IV) Actual procedures'''
- (April 3) Made concentrated top agar stock and LB solution, autoclaved and should be ready for dilution/use
- (April 3) Made concentrated top agar stock and LB solution, autoclaved and should be ready for dilution/use
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{| class="wikitable"
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|+LB and Top Agar Recipes (x8)
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|-
 +
|Content
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|Pure LB
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|Top Agar (3.2% stock)
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|-
 +
|dH20
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|300mL
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|250mL
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|-
 +
|NaCL
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|3.01g
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|2.04g
 +
|-
 +
|Tryptone
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|2.97g
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|2.03g
 +
|-
 +
|Yeast extract
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|1.52g
 +
|1.01g
 +
|-
 +
|Agar
 +
|0g
 +
|8.01g
 +
|}

Latest revision as of 13:27, 9 September 2013


Small Phage March - April Notebook: Experiments



Small Phage
March-April
May-June
July-August
September-October

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.

3.2% stock: 60mL
2.4% stock: 120mL
1.6% stock: 100mL
0.8% stock: 200mL

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

LB and Top Agar Recipes (x8)
Content Pure LB Top Agar (3.2% stock)
dH20 300mL 250mL
NaCL 3.01g 2.04g
Tryptone 2.97g 2.03g
Yeast extract 1.52g 1.01g
Agar 0g 8.01g



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