Noi:LabNotes/2010-10-21: Difference between revisions
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>Noi (Created page with '==Probe prep for the DMR220k_S4 and S5== (followed Dr. Zhang's conditions) ===Production PCR=== 10nM 1st …') |
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===Production PCR=== | ===Production PCR=== | ||
10nM 1st round amplicon 0.2ul | |||
2x Kapa SYBG qPCR Master Mix 50ul | 2x Kapa SYBG qPCR Master Mix 50ul | ||
100uM pAP1V61U 0.4ul | 100uM pAP1V61U 0.4ul | ||
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95C 30sec -> (95C 3sec -> 55C 30sec-> 60C 20sec) x 16 -> 60C 2min -> 15C hold. | 95C 30sec -> (95C 3sec -> 55C 30sec-> 60C 20sec) x 16 -> 60C 2min -> 15C hold. | ||
Ethanol precipitation; re-purified with twelve Qiaquick columns, each eluted with 40ul EB. | Ethanol precipitation; re-purified with twelve Qiaquick columns, each eluted with 40ul EB (total vol: 12 x 40 = 480ul). | ||
DMR220k_S4 DMR220k_S5 | DMR220k_S4 DMR220k_S5 | ||
DNA concentration (ng/ul) | DNA concentration (ng/ul) | ||
O.D.260/280 | O.D.260/280 | ||
Yield: concxtotal vol. | Yield: concxtotal vol. |
Revision as of 14:51, 22 October 2010
Probe prep for the DMR220k_S4 and S5
(followed Dr. Zhang's conditions)
Production PCR
10nM 1st round amplicon 0.2ul 2x Kapa SYBG qPCR Master Mix 50ul 100uM pAP1V61U 0.4ul 100uM AP2V6 0.4ul H2O 49ul =================================== 100ul x 96
95C 30sec -> (95C 3sec -> 55C 30sec-> 60C 20sec) x 16 -> 60C 2min -> 15C hold. Ethanol precipitation; re-purified with twelve Qiaquick columns, each eluted with 40ul EB (total vol: 12 x 40 = 480ul).
DMR220k_S4 DMR220k_S5 DNA concentration (ng/ul) O.D.260/280 Yield: concxtotal vol.