We have had some difficulty when running GBS pools on the Pippin Prep. The pools they produce are at times showing a lack of abundance. We are running this experiment to gain insight on the following questions:
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Pull ALF_GBS_10-11 PCR A Pool and add 100uL of TE to the pool to ensure there is enough sample to run on 8 lanes. (Pippin Prep recommends bringing DNA up to 30uL using TE, so this should not interfere).
Run Pippin Prep GBS protocol * range 300-450bp with Marker L with the lanes as follows:
Lane 5: Control
Lane 4: ALF_GBS R4 (Replicate 4)
Lane 3: ALF_GBS R3
Lane 2: ALF_GBS R2
Land 1: ALF_GBS R1
Program GBS protocol on Blue Pippin and run in the same structure as listed above . with Marker M1
Run all 8 replicates on the BioAnalyzer
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