Noi:LabNotes/2010-10-21
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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.
DMR220k_S4 DMR220k_S5 DNA concentration (ng/ul) O.D.260/280 Yield: concxtotal vol.