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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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sig70-F3 | 1.62 | 1.86 | 41323 | 16529.2 | 1192.46 | 3.4 | 46.6
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sig70-F4 | 1.70 | 1.99 | 32365 | 12946 | 933.96 | 4.3 | 45.7
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sig70-F5 | 1.72 | 2.07 | 33688 | 13475.2 | 972.14 | 45.9
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sig70-F6 | 2.07 | 1.95 | 45412 | 18164.8 | 1310.46 | 3.1 | 46.9
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sig70-F7 | 1.64 | 1.95 | 39046 | 15618.4 | 1126.76 | 3.6 | 46.4
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sig70-F8 | 1.43 | 1.63 | 45745 | 18298 | 1320.07 | 3.0 | 47.0
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sig70-F9 | 1.75 | 2.16 | 23628 | 9451.2 | 681.84 | 5.9 | 44.1
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sig70-F10 | 1.72 | 2.01 | 32682 | 13072.8 | 943.11 | 4.2 | 45.8
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sig70-F11 | 1.71 | 2.07 | 31494 | 12597.6 | 908.83 | 4.4 | 45.6
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sig70-F12 | 1.59 | 1.82 | 41202 | 16480.8 | 1188.97 | 3.4 | 46.6
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sig70-F13 | 1.73 | 2.03 | 29224 | 11689.6 | 843.3 | 4.7 | 45.3
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sig70-F14 | 1.41 | 1.58 | 45645 | 18258 | 1317.2 | 3.0 | 47.0
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sig70-F15 | 1.59 | 1.85 | 40681 | 16272.4 | 1173.9 | 3.4 | 46.6
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sig70-F16 | 1.57 | 1.82 | 42241 | 16896.4 | 1219.0 | 3.3 | 46.7
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sig70-R1 | 2.06 | 2.17 | 7516 | 3006.4 | 216.9 | 18.4 | 31.6
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sig70-R2 | 2.04 | 2.25 | 15255 | 6102.0 | 440.2 | 9.1 | 40.9
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sig70-R3 | 1.76 | 1.85 | 42503 | 17001.2 | 1226.5 | 3.3 | 46.7
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sig70-R4 | 2.01 | 2.21 | 24012 | 9604.8 | 692.9 | 5.8 | 44.2
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sig70-R5 | 2.10 | 2.27 | 18530 | 7412 | 534.7 | 7.5 | 42.5
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sig70-R6 | 1.98 | 2.19 | 28452 | 11380.8 | 821.0 | 4.9 | 45.1
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sig70-R7 | 1.94 | 2.17 | 32976 | 13190.4 | 951.6 | 4.2 | 45.8
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sig70-R8 | 2.00 | 2.20 | 25379 | 10151.6 | 732.4 | 5.5 | 44.5
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sig70-R9 | 2.00 | 2.24 | 24141 | 9656.4 | 696.6 | 5.9 | 44.1
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sig70-R10 | 1.94 | 2.17 | 30237 | 12094.8 | 872.6 | 4.6 | 45.4
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sig70-R11 | 1.92 | 2.16 | 31497 | 12598.8 | 908.9 | 4.4 | 45.6
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sig70-R12 | 1.92 | 2.17 | 30045 | 12018 | 867.0 | 4.6 | 45.4
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sig70-R13 | 2.01 | 2.18 | 18726 | 7490.4 | 540.4 | 7.4 | 42.6
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sig70-R14 | 1.88 | 2.03 | 36111 | 14444.4 | 1042.1 | 3.8 | 46.2
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sig70-R15 | 1.98 | 2.17 | 10679 | 4271.6 | 308.2 | 13 | 37
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sig70-R16 | 1.86 | 2.01 | 35969 | 14387.6 | 1038 | 3.9 | 46.1
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