Content Comparison

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•  Add 5 ul to each rxn
•  Pipette mix Library Prep Mix A 15X at ~130 ul and place on ice
•  Add 20 ul Lib Prep Mix A to each sample (viscous)
•  Pipette Mix 10X at 40 ul, brief centrifuge to collect
•  Incubate 20 C for 20 minutes (no heated lid)
•  Prepare at least 400 ul 85% EtOH per sample
•  Resuspend Illumina Purification Beads by vortex < 30s
•  Remove ligation rxn from incubation
•  Add 60 ul Illumina Purification Beads to each sample, pipette mix
•  Incubate 5 minutes at RT
•  Place on magnet 5 minutes
•  Remove ~135 ul supernatant
•  Add 200 ul 85% EtOH, incubate 30 s, and discard
•  Add 200 ul 85% EtOH, incubate 30 s, and discard
•  Pipette again to remove all EtOH
•  Air Dry for 2-5 minutes
•  Remove from magnet and add 34 ul ultra pure water, Pipette mix >10X at 32.5 ul
•  Incubate 5 minutes at RT
•  Return tubes to magnet for 2 minutes
•  Transfer 32.5 ul to new 0.2 ml tubes, store on ice
•  Thaw 4X PCR MM and UDI Index Mix strip
•  Add 5 ul Unique UDI Library Index Mix to each tube and record
•  Add 12.5 ul 4X PCR MM
•  Pipette Mix 10X at 32ul
•  Thermocycle with lid at 105C:

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9 samples to reamplify because of vortexing error: Wells B9, C9, A11, B11, C11, D11, H11, A12, and B12

Run on Thermocycler Illumina ReAmp:

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