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==== <u>1X AMPure bead purification</u> ==== * The instruction suggested to use 0.8X of AMPure bead for clean up, but I use 1X of AMPure bead - Aliquot AMPure bead in 1.5mL tube and leave at RT :- Add 55ul AMPure (RT) bead Mix by pipetting 10x :- Incubate at RT for 15min :- Transfer to sit on magnet for 5min :- Discard spnt :- Wash twice with 200ul freshly prepared 80% EtOH ::- After adding 80% EtOH, wait for 30sec (only the 1st time wash) before pipetting up and down 5X and discard spnt :- Dry the bead for 5min :- Resuspend with 21ul EB buffer :- Transfer 20ul of eluted adaptor-ligated DNA to new 8-tube strip ::- Before continue to bisulfite conversion, we can check if ligation works by running PCR using 1ul of adaptor-ligated DNA in small volume. (I skipped this step) :- Continue to bisulfite conversion. Optional: Adaptor-ligated DNA can be stored at -20C before bisulfite conversion
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