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.
Bruce Roe, broe@ou.edu