Yu:homemade taq

Colony PCR

Reagent	1.5x	2.5x	3.5x	6.5x	9.5x	12.5x
Fwd. Primer 10 μM	1.5 μL	2.5 μL	3.5 μL	6.5 μL	9.5 μL	12.5 μL
Rev. Primer 10 μM	1.5 μL	2.5 μL	3.5 μL	6.5 μL	9.5 μL	12.5 μL
10 mM ea dNTPs	1.5 μL	2.5 μL	3.5 μL	6.5 μL	9.5 μL	12.5 μL
DNA Template	5 μL of Lyse'n Go per reaction; Lyse'n Go thermocycle takes ~18 min. Can start Lyse'n Go cycle before making master mix
10x Rxn Buffer (Invitrogen (-MgCl2))	7.5 μL	12.5 μL	17.75 μL	32.5 μL	47.5 μL	62.5 μL
50 mM MgCl2	2.25 μL	3.75 μL	5.25 μL	9.75 μL	14.25 μL	18.75 μL
milliQ H2O	45.75 μL	76.25 μL	106.75 μL	198.25 μL	289.75 μL	381.25 μL
Homemade Taq	7.5 μL	12.5 μL	17.5 μL	32.5 μL	47.5 μL	62.5 μL
Add 45 μL of reaction master mix to each Lyse'n Go reaction