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
|
Ai5043 |
did not amplify |
did not amplify; remove from multiplex
|
Acos3 |
amplified well; no changes needed |
still fine, though seems to decrease with increasing Acos5 & Ai5071
|
Acos5 |
amplified with high intensity, pull down a bit |
0.04 = too low; 0.06 = inconclusive (failures); 0.08 = too high
|
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
|
Ai5071 |
amplified with lower intensity; increase a bit |
best at 0.24, 0.28, 0.32 (200-300 intensity only)
|
- 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
|
Ai5033 |
amplified well; no changes needed |
still fine (no amp for any markers in Ai5057 assay)
|
Ai5057 |
did not amplify |
did not amplify; remove from multiplex
|
Ai5037 |
amplified well; no changes needed |
still fine (no amp for any markers in Ai5057 assay)
|
Ai5035 |
amplified well; no changes needed |
a bit low in Ai5062 assay (no amp for any markers in Ai5057 assay)
|
Ai5062 |
amplified but with very low intensity; increase concentration |
best at 0.48 (400 & 6000 intensity)
|
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