ET Terminator Reactions
Written by Linda Ray
Updated 04-01-00

Here is the protocol: Not hard at all.
We need to order more mix before people can try this.


Reactions:

		full rxn:
		                    2 ul DNA (standard double acetate protocol) 0.2-2 ug
		                    1 ul primer 6.5 umoles
		                    8 ul ET premix
		                    q.s. dd H20
		                    20 ul

		1/2 rxn:
		                    2 ul DNA (standard double acetate protocol) 0.2-2 ug
		                    1 ul primer 6.5 umoles
		                    4 ul ET premix
		                    q.s. ddH20
		                    10 ul

		1/4 rxn:
		                    2ul DNA (standard double acetate protocol) 0.2-2 ug
		                    1ul primer 6.5 umoles
		                    2ul ET premix
		                    q.s. ddH20
		                    5-7 ul

Cyclying Conditions:

			Amersham recommends NOT to preheat thermocycler at 95 deg C but using
			a preheat has not been tested.

		        60 cycles of:
		
		        	95 deg C, 20 secs
		        	50 deg C, 15 secs
		        	60 deg C, 1 min
			
Post Processing:

			Filter through Sephadex G-50, store dry and then dissolve in water
			prior to loading as usual.
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Bruce Roe, broe@ou.edu