Matt Gethers/CRI, Thailand/Labwork/PCRs/PCR Pfu Template
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PCR PFU Template
Rxn Mix
| Reagent | Volume/Amount |
| Paeru Genomic DNA Template | 0.5 μl from So Pa Pan stock |
| Pfu | 0.5 μl |
| 4 μl 1 μM Stock | |
| 4 μl 1 μM Stock | |
| dNTPs | 1 μl 10 mM Stock |
| DMSO | 2.5 μl |
| Buffer S | 5 μl |
| dH20 | 32.5 μl |
| Total | 50 μl |
Rxn Conditions
Annealing Temperature: oC
Extension Time:
Cycle
| Step | Temperature | Duration | Notes |
| Initial Denaturation | 95oC | 3 minutes | |
| Repeat Cyclic Steps 35x | |||
| Cyclic Denaturation | 94oC | 40 Seconds | |
| Cyclic Annealing | oC | 40 Seconds | |
| Cyclic Extension | 72oC | minutes | |
| Repeat Cyclic Steps 35x | |||
| Final Extension | 72oC | 10 minutes |
