Not sure I’m going to get much more out of this flow cell, but I want to try one more wash and reload. Paused run at 2 days in. On this last wash/reload, I want to try getting pore occupancy higher, so I’ll try 3X the recommended concentration – 300fmol in 12uL EB. Since the pores are nearly exhausted, it’s lower stakes to try this now, in case overloading burns out the pores. This means I again need to increase the concentration of my MinION dilution of Library 4.
I want 300fmol = 129.3ng DNA in 12uL EB
I currently have the following:
undiluted G1L4: 3uL of 36.6ng/uL, so 109ng DNA
MinION dilution: 7ul of 7.18ng/uL, so 50.26ng DNA
To get 129.3ng DNA in 12uL EB: \((y*36.6\frac{ng}{uL}) + (7uL*7.18\frac{ng}{uL}) = 129.3ng \ \ \Rightarrow \ \ y = 2.16uL\)
So I need to combine:
7uL of MinION dilution = 50.26ng DNA
2.16uL of G1L4 = 79.05ng DNA
2.84uL EB
For a total of 129.31ng DNA in 12uL of EB.
Combined the above quantities to obtain 300fmol in 12uL EB. then washed and reloaded the flow cell with this, and resumed run.
Before wash, 420 pores (128 reserved, 292 available), after wash thre were 475 pores (208 reserved, 267 available). So no appreciable difference.