Additional instructions for Expand (Roche/Boehringer-Mannheim) Long Template PCR Master Mix I Master Mix II template enzyme primers buffer dNTPs ddH2O to 25 l ddH2O to 25 l Notes: Preheat PCR machine to 94°C using instant key Mix the above two for 1 reaction F.V. 50 l Use thin wall 0.2 ml PCR tubes Vortex to mix thoroughly Spin pulse Overlay with 30 l oil even with hot bonnet Master mix I can be mixed in thin wall tube when few samples Master mixes of I or II can be done for large # of reactions