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CAROL
Diluting Primers and Primer Extension Polymerase Chain Reaction (PCR)
- To create the synthetic sigma 70 promoter library, we have synthesized 32 different primers (16 different forward primers and 16 different reverse primers) to hopefully form 256 different promoters.
- Need to dilute primers to 80μM. The following is the concentration of the original primers that were synthesized once water was added to the primers.
Primer | 260/280 | 260/230 | Concentration [ng/μL] | Reading/2.5 [ng/μL] | Stock Concentration [μM] | Volume stock into working [μL] | Volume water into working [μL] |
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sig70-F1 | 1.98 | 1.73 | 20816 | 832.64 | 60.07 | 6.7 | 43.3
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sig70-F2 | 1.55 | 1.80 | 44284 | 17713.6 | 1277.91 | 3.1 | 46.9
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psB1AC3 3 | 1.87 | 1.88 | 48.5
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psB1AC3 4 | 1.90 | 2.11 | 118.9
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psB1AK3 1 | 2.05 | 2.42 | 134.4
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psB1AK3 2 | 1.95 | 2.23 | 55.6
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psB1AK3 3 | 1.91 | 2.22 | 97.9
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psB1AK3 4 | 2.03 | 1.92 | 44.5
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CHINMOYEE
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