Nuclear Region 454 Amplification
Oleg Samples Amplified with primer no adapters
Extend with primers w/adapters
| EX Takara RXN Mixture
|
conc
|
1X
|
8
|
'
|
| Milli Q water |
|
6.15 |
49.2 |
|
| Takara EX Buffer |
10X |
1 |
8 |
|
| dNTP |
2.5uM |
0.8 |
6.4 |
|
| Primer Mix |
20uM |
1 |
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| EX Taq |
5U |
0.05 |
0.4 |
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| Template |
Oleg PCR 1:10 |
1 |
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64 |
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8.0ul / RXN add 1.0ul Primer and 1.0ul PCR template |
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| PCR cycle |
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| 98C |
10 sec |
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| 60C |
1 min |
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| x5 |
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| AMP Lane |
Template |
Primer |
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| 1 |
Culture 11 |
B11 |
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| 2 |
Culture 11 |
D11 |
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| 3 |
Culture 11 |
E11 |
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| 4 |
Culture 11 |
F11 |
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| 5 |
Culture H |
B12 |
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| 6 |
Culture H |
C12 |
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| 7 |
Culture H |
E12 |
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| 8 |
Culture H |
F12 |
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Load 2.0ul of sample
Placed same PCR on machine for an additional 15 cycles
Load 2.0ul of sample
|
Alien genes
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