Pippin Prep vs Blue Pippin Experiment

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:

1. Does the Blue Pippin produce more bountiful elutions than the Pippin Prep in the 300-450bp range?

2. Are all lanes comparable on the Pippin Prep when it comes to quality elutions?

3. Are lanes comparable between the Pippin Prep and Blue Pippin?

Protocol

• 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

Results

• ALF_GBS PP vs BP Gel:

• ALF_GBS PP EPG:

• ALF_GBS BP EPG:

• ALF_GBS PP vs BP R1 EPG:

• ALF_GBS PP vs BP R2 EPG:

• ALF_GBS PP vs BP R3 EPG:

• ALF_GBS PP vs BP R4 EPG: