Team:University of Lethbridge/Notebook/GeneralLabAugst

From 2008.igem.org

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Tranformation of pUC19:
Tranformation of pUC19:
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1. Thaw E.coli DH5a cells on ice
+
1. Thaw E.coli DH5a cells on ice
-
2. Add pUC19 DNA, pipette gently to mix (1μl of plasmid)
+
2. Add pUC19 DNA, pipette gently to mix (1μl of plasmid)
-
3. Let sit for 30 minutes on ice
+
3. Let sit for 30 minutes on ice
-
4. Incubate cells for 30 seconds at 42oC
+
4. Incubate cells for 30 seconds at 42oC
-
5. Incubate cells on ice for 2 min
+
5. Incubate cells on ice for 2 min
-
6. Add 1 mL SOC  at room temp
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6. Add 1 mL SOC  at room temp
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7. Incubate for 1 hour at 37oC on shaker
+
7. Incubate for 1 hour at 37oC on shaker
-
8. Spread 100-300 μl onto a plate made with appropriate antibiotic
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8. Spread 100-300 μl onto a plate made with appropriate antibiotic
-
9. Grow overnight at 37 °C
+
9. Grow overnight at 37 °C

Revision as of 02:37, 4 August 2008

August 03, 2008

Nathan Phillips

Tranformation of pUC19: 

1. Thaw E.coli DH5a cells on ice
2. Add pUC19 DNA, pipette gently to mix (1μl of plasmid)
3. Let sit for 30 minutes on ice
4. Incubate cells for 30 seconds at 42oC
5. Incubate cells on ice for 2 min
6. Add 1 mL SOC  at room temp
7. Incubate for 1 hour at 37oC on shaker
8. Spread 100-300 μl onto a plate made with appropriate antibiotic
9. Grow overnight at 37 °C
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