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CNHY MP1a/b Optimization
- optimize for marker peak height - vary each primer (based on results from 23 July 2012)
- CNHY MP1a: increase Ai5013, Ai5043, Ai5071
- CNHY MP1a: decrease Acos5
- CNHY MP1b: increase Ai5057, Ai5062
Frag Results
MP1a
| | 23 July 2012 | this time
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| Ai5043 | did not amplify | did not amplify; remove from multiplex
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| Acos3 | amplified well; no changes needed | still fine, though seems to decrease with increasing Acos5 & Ai5071
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| Acos5 | amplified with high intensity, pull down a bit | 0.04 = too low; 0.06 = inconclusive (failures); 0.08 = too high
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| Ai5013 | amplified with very low intensity; increase concentration | second peak never got above 250, regardless of concentration; but in Acos5 assay, noticed second peak reached 400 intensity as Acos5 decreased
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| Ai5071 | amplified with lower intensity; increase a bit | best at 0.24, 0.28, 0.32 (200-300 intensity only)
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- since removing the only marker labelled with FAM, move one of the NEDs to FAM
- NEDs: Acos5 and Ai5013 ... since Ai5013 tends to be low intensity, would be a good one to move to brighter FAM
MP1b
| | 23 July 2012 | this time
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| Ai5033 | amplified well; no changes needed | still fine (no amp for any markers in Ai5057 assay)
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| Ai5057 | did not amplify | did not amplify; remove from multiplex
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| Ai5037 | amplified well; no changes needed | still fine (no amp for any markers in Ai5057 assay)
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| Ai5035 | amplified well; no changes needed | a bit low in Ai5062 assay (no amp for any markers in Ai5057 assay)
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| Ai5062 | amplified but with very low intensity; increase concentration | best at 0.48 (400 & 6000 intensity)
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Tara's Lab Notebook
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