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Pool qPCR results:
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iSeq Run:
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
1000/32.25 = 31uL of Pool to Add
100 - uL of Pool to Add = ul of “10 mM Tris 8.5” to Add
1000- 31 = 969uL of 10mM Tris 8.5
*1nM pool will be combined with 1nM pool for the low read 2.5 experiment. Directions for iSeq prep found here.