...
Add 6 ul 1 nM Pool to 82 ul “10 mM Tris 8.5” and 12 ul 50 pM PhiX
Remove iSeq 100 i1 Flow Cell from refrigerator 5’s crisper drawer and open white foil pack and allow to equilibrate to RT for 10-15 minutes.
Open “iSeq 100 i1 Reagent Cartridge v2”. Turn on iSeq100
Click on “Sequence”. Watch Video. Do what video tells you to do. Follow on screen instructions until run starts.
Redone Sequencing Test:
Dilute to 1 nM based off qPCR results. qPCR results are in pM, but 1:1000 dilution used. The results are effectively in nM for pool.
1000/Results = ul of Pool to Add
100/10 = 10uL of Pool to Add
100 - uL of Pool to Add = ul of “10 mM Tris 8.5” to Add
100- 10 = 90 uL of 10mM Tris 8.5
Dilute 1 nM full pool to loading concentration of 60 pM:
Add 6 ul 1 nM Pool to 82 ul “10 mM Tris 8.5” and 12 ul 50 pM PhiX
Remove iSeq 100 i1 Flow Cell from refrigerator 5’s crisper drawer and open white foil pack and allow to equilibrate to RT for 10-15 minutes.
Open “iSeq 100 i1 Reagent Cartridge v2”. Turn on iSeq100
Click on “Sequence”. Watch Video. Do what video tells you to do. Follow on screen instructions until run starts.