Latest revision as of 14:35, 26 October 2010

Growing an Overnight Culture

Transfer 5 ml of LB or appropriate media into a universal tube.
Heat the innoculation loop till red hot to sterilize the loop.
Allow the innoculation loop to cool down for approximately 30 seconds.
The innoculation loop is then used to pick a single colony of bacteria on the agar plate.
Innoculate the LB or appropriate media with the bacteria.
Resterilize the loop.
The culture is then left overnight at 37°C in a shaking incubator at 200 rpm.

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 ==Procedures==
-# 5ml of LB broth is pipetted with the help of an auto pipette into a universal tube.
+# Transfer 5 ml of LB or appropriate media into a universal tube.
-# Heat the flame loop with the help of a burner till the loop is red hot to kill all the bacteria present on the loop.
+# Heat the innoculation loop till red hot to sterilize the loop.
-# Let the flame loop to cool down for approximately 2 minutes.
+# Allow the innoculation loop to cool down for approximately 30 seconds.
-# The flame loop is then used to pick a colony of the bacteria present on the agar plate.
+# The innoculation loop is then used to pick a single colony of bacteria on the agar plate.
-# The colony is then mixed into the LB Broth in the universal tube.
+# Innoculate the LB or appropriate media with the bacteria.
-# Again flame the flame loop till it gets red hot so as to kill any bacteria present on it.
+# Resterilize the loop.
-# The culture is then left overnight at 37°C in an incubator.
+# The culture is then left overnight at 37°C in a shaking incubator at 200 rpm.