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

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>Alan6017518
 
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==Quantification of New amplified Phi-X and all libraries==
==Quantification of New amplified Phi-X and all libraries==
*Dilute concentration by 10 fold for Har_2, Har_5
*Dilute concentration by 10 fold for Har_2, Har_5
*Dilute New amplified PhiX to 1nM
*Dilute New amplified PhiX to 1nM (Mix 2ul amp. PhiX with 164.97 ddH2O)
*Keep P3, P5, P7 and CVF_N_D as is  
*Keep P3, P5, P7 and CVF_N_D as is  
{| {{table}}
{| {{table}}
Line 42: Line 42:
| H20||24.000||192||211.2
| H20||24.000||192||211.2
|-
|-
|
|}
{| {{table}}
| ||nM||sample ul||Tris-CL ul||HP3||total
|-
| 8/9 P7||8.09||3||20.06||1.21||24.27
|-
| CVF_N_D||2.91||4||7.06||0.58||11.64
|-
|
|}
{| {{table}}
| Dilute to 10nM||Sample (ul)||||ddH2O
|-
| 8/9 P3||4.000||6.928||2.928
|-
| 8/9 P5||4.000||6.628||2.628
|-
| Har_2||4.000||5.600||1.600
|-
| Har_5||4.000||6.876||2.876
|-
| CVI||5||-||18.546
|-
|CVF||5||-||4.710
|  
|  
|}
|}

Latest revision as of 18:20, 23 November 2009

Quantification of New amplified Phi-X and all libraries[edit]

  • Dilute concentration by 10 fold for Har_2, Har_5
  • Dilute New amplified PhiX to 1nM (Mix 2ul amp. PhiX with 164.97 ddH2O)
  • 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
nM sample ul Tris-CL ul HP3 total
8/9 P7 8.09 3 20.06 1.21 24.27
CVF_N_D 2.91 4 7.06 0.58 11.64
Dilute to 10nM Sample (ul) ddH2O
8/9 P3 4.000 6.928 2.928
8/9 P5 4.000 6.628 2.628
Har_2 4.000 5.600 1.600
Har_5 4.000 6.876 2.876
CVI 5 - 18.546
CVF 5 - 4.710