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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==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 28: | Line 28: | ||
{| {{table}} | {| {{table}} | ||
| |||| | | ||||X8||X1.1 | ||
|- | |- | ||
| PhiX Control DNA (10nM)||1.000|||| | | PhiX Control DNA (10nM)||1.000|||| | ||
|- | |- | ||
| Syb_FP5 (100uM)||0.200|| | | Syb_FP5 (100uM)||0.200||1.6||1.76 | ||
|- | |- | ||
| Syb_RP7 (100uM)||0.200|| | | Syb_RP7 (100uM)||0.200||1.6||1.76 | ||
|- | |- | ||
| 2X Phusion||25.000|| | | 2X Phusion||25.000||200||220 | ||
|- | |- | ||
| 50X SYBR Green I||0.200|| | | 50X SYBR Green I||0.200||1.6||1.76 | ||
|- | |- | ||
| H20||24.000|| | | 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 |