We are expecting little to no difference. So, we will proceed with actual template, but the smaller PCR pool. We will compare via qPCR and bioanalyzer.
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Template | EcoR1 MID plate |
|---|---|
2017_ALFALFA_PLATE10 | GBS EcoR1 Plate 5 |
2017_ALFALFA_PLATE11 | GBS EcoR1 Plate 6 |
Label reaction plates with MID plate used.
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Create the two pools below using 20uL from ligated plates matching well to well.
Pool | Plate1 | Plate2 | Plate3 | Plate4 |
|---|---|---|---|---|
2A | 2017_ALFALFA_PLATE10_Thermocycler | 2017_ALFALFA_PLATE11_Thermocycler |
|
|
2B | 2017_ALFALFA_PLATE10_Incubator | 2017_ALFALFA_PLATE11_Incubator |
...
GBS10-11 PCR_A Col_1: 78.13 nanomoles (thermocycler)
GBS10-11 PCR_A Col_6: 85.33 nanomoles (thermocycler)
GBS10-11 PCR_A Col_12: 75.57 nanomoles (thermocycler)
GBS10-11 PCR_B Col_1: 96.37 nanomoles (incubator)
GBS10-11 PCR_B Col_6: 78.72 nanomoles (incubator)
GBS10-11 PCR_B Col_12: 80.08 nanomoles (incubator)
GBS10-11 PCR_A Pool: 95.64 nanomoles (thermocycler)
GBS10-11 PCR_B Pool: 110.7 nanomoles (incubator)
GBS10-11 Pool: 95.91 nanomoles
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