Team:Caltech/Protocols

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Caltech iGEM 2011

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Recipe for Agar/LB plate:

1) Fill two bottles with 500 g of nanopure water, 10 g of Tryptone, 10 g of NaCl, 5 g of yeast extract, and 15 g of Agar.

2) Shake and then autoclave.

Recipe for 50% Glycerol Stock: 1) Pour 50 mL of Glycerol and 50 mL of pure water into a graduated cylinder. 2) Cover with parafilm and turn over a couple of times to mix. 3) Use the Nalgene vacuum to sterilize the stock. 4) Use the bunsen burner to keep air sterile and close the cap.

Recipe for Enrichment Minimal Media consists of

1.0712 g of K2 HPO4
0.5239 g of KH2 PO4
79 mL of Phosphate solution
100 mL of Salt Solution I
100 mL of Salt Solution II
10 mL of Wolfe's Vitamin Solution
1 mL of SL-10 Trace Element Solution.

Transforming DNA Protocol: 1) Thaw competent cells on ice: 15K, 15J, 15O, 15M. 2) Get 10 micro Liters of pure water. 3) Pipette out DNA from the source plate and put it in the appropriate tubes. 4) Flick the competent cells in the tube gently. 5) Pipette 40 micro Liters of competent cells into the DNA. 6) Pipette 2 micro Liters of the DNA and put it in the competent cell tubes. 7) Stir a but. 8) Leave on ice for 30 minutes. 9) Heat Shock by using a water bath set to 42°C.

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 {{Team:Caltech/templateheader|
 Content=
+<p>Recipe for Agar/LB plate:
+) Fill two bottles with 500 g of nanopure water, 10 g of Tryptone, 10 g of NaCl, 5 g of yeast extract, and 15 g of Agar.
+) Shake and then autoclave.</p>
+<p>Recipe for 50% Glycerol Stock:
+) Pour 50 mL of Glycerol and 50 mL of pure water into a graduated cylinder.
+) Cover with parafilm and turn over a couple of times to mix.
+) Use the Nalgene vacuum to sterilize the stock.
+) Use the bunsen burner to keep air sterile and close the cap.</p>
+Recipe for Enrichment Minimal Media consists of
+* 1.0712 g of K2 HPO4
+* 0.5239 g of KH2 PO4
+* 79 mL of Phosphate solution
+* 100 mL of Salt Solution I
+* 100 mL of Salt Solution II
+* 10 mL of Wolfe's Vitamin Solution
+* 1 mL of SL-10 Trace Element Solution.
+Transforming DNA Protocol:
+) Thaw competent cells on ice: 15K, 15J, 15O, 15M.
+) Get 10 micro Liters of pure water.
+) Pipette out DNA from the source plate and put it in the appropriate tubes.
+) Flick the competent cells in the tube gently.
+) Pipette 40 micro Liters of competent cells into the DNA.
+) Pipette 2 micro Liters of the DNA and put it in the competent cell tubes.
+) Stir a but.
+) Leave on ice for 30 minutes.
+) Heat Shock by using a water bath set to 42°C.
 }}