From 2008.igem.org

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 {{:Team:Newcastle University/Header}}
-{{:Team:Newcastle University/Template:UnderTheHome|page-title=[[Team:Newcastle University/Protocols|Protocols]]}}
+{{:Team:Newcastle University/Template:UnderTheWetLab|page-title=[[Team:Newcastle University/Protocols|Protocols]]}}
+{{:Team:Newcastle University/WetlabSideBar}}
+<div id="maincol">
+==Protocols==
+[[Agarose Gel Electrophoresis]]
-== Agarose Gel Electrophoresis ==
+[[Making Agar Plates]]
-* Add 20ml 50 X TAE to 980ml H2O.
+[[Isolating Plasmid from Cells (Miniprep)]]
-* Add 3g agarose powder to 300ml of the TAE-H2O (1 x TAE) solution.
-* Microwave full power for 5 minutes or until solution is clear and agarose has dissolved.
-* Leave to stand for 5-10 minutes. Put tape around sides of tray.
-*Pour solution into tray, add 4ul of ethidium bromide and mix gently using the comb.
-* Clean the comb (this prevents capillary action drawing the gel up between the comb teeth and making 'spikes' around the wells) and reinsert near one end of the tray.
-* Leave to set 30-40 mins.
-* Remove tape and comb, place tray in electrophoresis machine and pour 1 x TAE solution to just cover the gel.
-* Combine 1ul of loading buffer for every 5ul sample being loaded and mix by pipetting up and down.
-* Load marker and samples and run the gel.
-N.B. We have found that a thin gel works best for loading small samples, but a thicker gel may be preferable if lots of sample needs to be loaded (for example if a fragment needs to be cut from the gel).
+[[Restricting Plasmids (Double Restriction)]]
-N.B. We have run gels at 100V for 20mins and 70V for 60mins. The latter setting/time is better for running larger fragments such as whole plasmids as a high voltage can cause shearing of the DNA.
+[[Purifying DNA from an enzymatic reaction]]
+[[Cutting a Specific Band from Agarose Gel]]
-== Isolating Plasmid from Cells (Miniprep) ==
+[[Purifying DNA from Gel Slices]]
+[[Ligating DNA]]
+[[Transforming into DH5α or TOP10 E. coli]]
+[[Making Frozen Glycerol Cell Stocks From Overnight Cultures]]
+[[Making Overnight Cultures from Frozen Glycerol Cell Stocks]]
+[[Making Overnight Cultures from Agar Colonies]]
+[[Transforming into Bacillus subtillis]]
+[[Inducing Bacillus subtilis with Subtilin]]
+[[Preparing Bacillus subtilis for Microscopy]]
+</div>