AlanFung:LabNotes/Sequencing/2009-11-23: Difference between revisions

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>Alan6017518
(New page: ==Quantification of New amplified Phi-X and all libraries== *Dilute concentration by 10 fold for Har_2, Har_5 *Dilute New amplified PhiX to 1nM *Keep P3, P5, P7 and CVF_N_D as is {| {{tab...)
 
>Alan6017518
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{| {{table}}
{| {{table}}
| ||||X20||X1.1
| ||||X8||X1.1
|-
|-
| PhiX Control DNA (10nM)||1.000||||
| PhiX Control DNA (10nM)||1.000||||
|-
|-
| Syb_FP5 (100uM)||0.200||4.000||4.400
| Syb_FP5 (100uM)||0.200||1.6||1.76
|-
|-
| Syb_RP7 (100uM)||0.200||4.000||4.400
| Syb_RP7 (100uM)||0.200||1.6||1.76
|-
|-
| 2X Phusion||25.000||500.000||550.000
| 2X Phusion||25.000||200||220
|-
|-
| 50X SYBR Green I||0.200||4.000||4.400
| 50X SYBR Green I||0.200||1.6||1.76
|-
|-
| H20||24.000||480.000||528.000
| H20||24.000||192||211.2
|-
|-
|  
|  
|}
|}

Revision as of 16:30, 23 November 2009

Quantification of New amplified Phi-X and all libraries

  • Dilute concentration by 10 fold for Har_2, Har_5
  • Dilute New amplified PhiX to 1nM
  • Keep P3, P5, P7 and CVF_N_D as is
Samples
PhiX-Stock 8/9 P7
PhiX-Stock 8/9 P7
PhiX(1nM) Har_2
PhiX(1nM) Har_2
8/9 P3 Har_5
8/9 P3 Har_5
8/9 P5 CVF_N_D
8/9 P5 CVF_N_D
  • split every 50ul rxn into 2 wells
98C 30 sec -> 20 cycles of (98C 10sec->64C 20sec->72C 20sec)->72C 5 min
X8 X1.1
PhiX Control DNA (10nM) 1.000
Syb_FP5 (100uM) 0.200 1.6 1.76
Syb_RP7 (100uM) 0.200 1.6 1.76
2X Phusion 25.000 200 220
50X SYBR Green I 0.200 1.6 1.76
H20 24.000 192 211.2