qPCR
Make 1:1000 dilutions of all samples using 2 ml tubes
Add 16 ul of Illumina Library Quantification MasterMix to each well:
ul/rxn | Reagent | # of rxns | ul needed |
|---|---|---|---|
10 ul | KAPA SYBR FAST qPCR MM (2X) | 40 | 400 |
2 ul | Primer Premix (10X) | 40 | 80 |
4 ul | Ultra Pure Water | 40 | 160 |
16 ul | Total Volume | 40 | 640 |
Add 4 ul of template, pool, or standards to each well:
| 1 | 2 | 3 | 4 | 5 | 6 | 7 | 8 | 9 | 10 | 11 | 12 |
|---|---|---|---|---|---|---|---|---|---|---|---|---|
A | 5pM PhiX | 5pM PhiX | 5pM PhiX |
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| NTC | NTC | NTC | ||
B | LRII.5 | LRII.5 | LRII.5 |
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| 0.0002 pM Std | 0.0002 pM Std | 0.0002 pM Std | ||
C | locad2 | locad2 | locad2 |
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| 0.002 pM Std | 0.002 pM Std | 0.002 pM Std | ||
D | 5pM PhiX | 5pM PhiX | 5pM PhiX |
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| 0.02 pM Std | 0.02 pM Std | 0.02 pM Std | ||
E |
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| 0.2 pM Std | 0.2 pM Std | 0.2 pM Std | |||||
F |
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| 2 pM Std | 2 pM Std | 2 pM Std | |||||
G |
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| 20 pM Std | 20 pM Std | 20 pM Std | |||||
H |
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