RAPD PCR Reaction Mixes:
| Mg++[uM]
| 1.5
| 2.0
| 2.5
| 3.0
| 3.5
|
| DNA Template [10 ng/ul]
| 2.0
| 2.0
| 2.0
| 2.0
| 2.0
|
| 10X PCR buffer
| 2.5
| 2.5
| 2.5
| 2.5
| 2.5
|
| MgCl2 [25mM]
| 1.5
| 2.0
| 2.5
| 3.0
| 3.5
|
| Primer [2.5uM]
| 2.0
| 2.0
| 2.0
| 2.0
| 2.0
|
| dNTP (10mM, 2.5mM each)
| 2.0
| 2.0
| 2.0
| 2.0
| 2.0
|
| H2O
| 14.9
| 14.4
| 13.9
| 13.4
| 12.9
|
| Taq Polymerase (0.5u/ul)
| 0.1
| 0.1
| 0.1
| 0.1
| 0.1
|
| Total Volume (ul)
| 25.0
| 25.0
| 25.0
| 25.0
| 25.0
|
PCR Reaction Conditions:
|
|
| Cycles
|
|
| Type
| Initialize
| X
| Denature
| Anneal
| Extend
| Termination
|
| RAPD 10-mer
| 95C, 5min
| 45
| 94C, 0:30
| 35C, 1:00
| 72C, 2:00
| 72C, 5:00
|
| PCR 18+mer
| 95C, 5:00
| 30
|
|
|
|
|
| PCR LMT
| 95C, 5:00
| 30
|
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