Nuclear Region 454 Amplification
Oleg Samples Amplified with primer no adapters
Extend with primers w/adapters
EX Takara RXN Mixture
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conc
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1X
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8
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'
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Milli Q water |
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6.15 |
49.2 |
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Takara EX Buffer |
10X |
1 |
8 |
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dNTP |
2.5uM |
0.8 |
6.4 |
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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
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