Setup Notes
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Normalize plate reader with Buffer AE for all plates
Quantify all plates
Check for samples above 150 ng/ul
Transfer 20 ul from all wells of a plate needing normalization to new VWR plate
Add 20 ul TE to all wells above 150 ng/ul
Restriction Digestion
(Keep MM and reaction plates on ice)
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Reagent | ul/rxn | rxns | ul needed (x1.4) | ||
|---|---|---|---|---|---|
H2O | 9.52 | 576 | 7712 | 5483.52 | 19045712 |
5x iProof buffer | 4 | 576 | 3240 | 23042400 | 800 |
10 mM dNTPs | 0.4 | 576 | 324 | 230.4 | 80240 |
50 mM MgCl2 | 0.4 | 576 | 324 | 230.4 | 80240 |
5 uM Illumina Primers | 1.33 | 576 | 1077 | 766.08 | 266798 |
iProof TAQ | 0.2 | 576 | 162 | 115.2 | 40120 |
DMSO | 0.15 | 576 | 121 | 86.490 | 30 |
total | 16 | 576 | 12960 | 9216 | 32009600 |
Add 16 ul of PCR1 MM to each well of four new hard shell PCR plates with 8 channel
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