AlanFung:LabNotes/Sequencing/2009-11-19: Difference between revisions

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>Alan6017518
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| H20||24.000||480.000||528.000
| H20||24.000||480.000||528.000
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{| {{table}}
| Result||||||||
|-
| PhiX||9.112||Cycle Difference||2^||x10 (nM)
|-
| Amp PhiX||6.051||3.062||8.348||83.484
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| CVI||6.877||2.236||4.709||47.093
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| CVF||8.155||0.958||1.942||19.419
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| 1-7 PGP1||8.346||0.766||1.701||17.011
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| 8 PGP1||6.510||2.603||6.073||60.734
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| 9 PGP1||7.244||1.868||3.650||36.503
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| N12||7.915||1.198||2.293||22.934
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| PGP8B||11.776||2.664||6.339||1.577
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Revision as of 22:09, 19 November 2009

Quantification of New amplified Phi-X and all libraries

  • Dilute all samples by 10 fold
Samples
PhiX-Stock CVF PGP4 PGP10 PGP8C
PhiX-Stock CVF PGP4 PGP10 PGP8C
PhiX-Stock 1-7 PGP1 PGP6 NA12878 PGP8D
PhiX-Stock 1-7 PGP1 PGP6 NA12878 PGP8D
PhiX 8 PGP1 PGP8 PGP8A PGP8E
PhiX 8 PGP1 PGP8 PGP8A PGP8E
CVI 9 PGP1 PGP9 PGP8B N12
CVI 9 PGP1 PGP9 PGP8B N12
  • split every 50ul rxn into 2 wells
98C 30 sec -> 20 cycles of (98C 10sec->64C 20sec->72C 20sec)->72C 5 min
X20 X1.1
PhiX Control DNA (10nM) 1.000
Syb_FP5 (100uM) 0.200 4.000 4.400
Syb_RP7 (100uM) 0.200 4.000 4.400
2X Phusion 25.000 500.000 550.000
50X SYBR Green I 0.200 4.000 4.400
H20 24.000 480.000 528.000
Result
PhiX 9.112 Cycle Difference 2^ x10 (nM)
Amp PhiX 6.051 3.062 8.348 83.484
CVI 6.877 2.236 4.709 47.093
CVF 8.155 0.958 1.942 19.419
1-7 PGP1 8.346 0.766 1.701 17.011
8 PGP1 6.510 2.603 6.073 60.734
9 PGP1 7.244 1.868 3.650 36.503
N12 7.915 1.198 2.293 22.934
PGP8B 11.776 2.664 6.339 1.577