1. gDNA isolation  
  2. Hackflex prep – Contains workflows for Tagmentation, barcoding and library ampification PCR, Purification and size selection, and DNA quantification. Relevant steps can be pulled for your particular experiment.
  3. Sequencing  
    • Pooling: Add your barcoded libraries together in an even ratio at a target concentration  
    • Pooling Samples Evenly
    • Pooling Samples Evenly if you have very high and very low concentrations
      • Note: The minimum concentration you can use depends on your sequencer. You can convert to Molarity from ng/ul using your avg. Fragment size.
    • Adding your amplified library to a tube using values calculated in the “pooling samples evenly” xls file. This is done using a python script that generates an html file you can open on an iPad – useful and worth it!
      • Copy the folder Pipette-Guide-96 into your private directory. Replace “volumes.csv” with the “output of pooling samples evenly.xls” and run “makehtml.py” in that subdirectory (from terminal). Open the new html file on a mobile device and tap through the wells as you pipet.