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AlanFung:LabNotes/Sequencing/2009-11-23
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==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 (Mix 2ul amp. PhiX with 164.97 ddH2O) *Keep P3, P5, P7 and CVF_N_D as is {| {{table}} | 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 {| {{table}} | ||||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 |- | |} {| {{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 | |}
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