Yu:homemade taq
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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 |