Toronto/Lab Protocols/Overnight

< Toronto | Lab Protocols(Difference between revisions)

Latest revision as of 03:38, 27 October 2007

Put 2.5 mL of LB Broth into a 15 mL tube. (Make sure to flame the broth)
Scrape off a single colony from a plate with a pipette tip and eject tip into the tube.
Add 2.5 μL of the appropriate antibiotics. (Check spreadsheet)
Incubate overnight.

Jump to [http://igem.skule.ca/lab/protocols/miniprep.htm BlueGenes]

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-== Miniprep ==
+== Overnight (O/N) Preparation: ==
-Small Scale Plasmid Preparation (1 – 2 hours)
-'''Overnight (O/N) Preparation:'''
 #Put 2.5 mL of LB Broth into a 15 mL tube. (Make sure to flame the broth)
@@ Line 10: / Line 6: @@
 #Add 2.5 μL of the appropriate antibiotics. (Check spreadsheet)
 #Incubate overnight.
-'''Miniprep Procedure:'''
-#Put 1.5 mL O/N into a 1.5 mL Eppendorf tube.
-#Centrifuge for 1 minute.
-#Aspirate the supernatant.
-#In order to increase the concentration of DNA, add the rest of the o/n into the same tube and centrifuge for 1 minute again.
-#Aspirate the supernatant and resuspend cells with 250 μL Resuspension Solution (red). Vortex until pellet dissolves.
-#Add 250 μL Lysis Solution and mix with pipette. (shake with hand)
-#Sit for 3-5 minutes.
-#Add 350 μL Neutralization Solution (blue) and invert tube with hand 4-6 times.
-#Centrifuge for 5 minutes.
-#Pour supernatant in a column tube.
-#Centrifuge for 1 minute and throw away supernatant.
-#Add 500 μL Wash Solution.
-#Centrifuge for 1 minute and throw away supernatant.
-#Add 500 μL Wash Solution.
-#Centrifuge for 1 minute and throw away supernatant.
-#Put blue column in a fresh centrifuge tube. (make sure to label)
-#Add 50 μL Elution Buffer.
-#Incubate for 2 minutes at room temperature, then centrifuge for 2 minutes.
-#Throw away blue column.
-#Place plasmid into the freezer.
 Jump to [http://igem.skule.ca/lab/protocols/miniprep.htm BlueGenes]