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=== Post oxBS PCR of control DNA with TruS_F/R primers=== * Reaction set up: {| class="wikitable" | align="center" style="background:#f0f0f0;"|'''Reagent''' | align="center" style="background:#f0f0f0;"|'''Concentration''' | align="center" style="background:#f0f0f0;"|'''Volume (uL)''' | align="center" style="background:#f0f0f0;"|'''Final concentration''' | align="center" style="background:#f0f0f0;"|'''8.5X volume''' |- | Water||NA||40.65||NA||345.53 |- | PfuTurboCx Buffer||10X||5||1X||42.5 |- | TruS_F||100 uM||0.15||0.3 uM||1.275 |- | TruS_R||100 uM||0.15||0.3 uM||1.275 |- | dNTP|| 10 mM||1.25||250 nM||10.625 |- | PfuTurboCx||2.5U/uL||0.8||1 U||6.8 |- | Template||NA||2||NA||--- |} * I didn't get the 5hmC control out from the PCR. This is a big problem because without the 5hmC control we cannot determine the effectiveness of oxidation. I only saw bands at ~400 bp which means the 5mC and/or C survived the whole process. * 2 uL template (post oxBS elution), 20 cycles: [[File:control_dna_v3_post_oxBS_pcr_2ul.jpg | 300px]] * 10 uL template (post oxBS elution), 15 cycles: [[File:control_dna_v3_post_oxBS_pcr_10ul.jpg | 300px]]
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