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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.

Templates:

ALF_PLT10_2017_GBS

ALF_PLT11_2017_GBS

Restriction Digestion

(Keep MM and reaction plates on ice)

...

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

...