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===Adapter ligation=== * 30uM Y-adapter made by Alice few weeks ago * 0.08ul for sample 1,3,4,5,7 and 0.4ul for 2,6,8 * 50ul total with 10ul buffer and 5ul Quick T4 ligase; RT for 30min {| {{table}} | align="center" style="background:#f0f0f0;"|'''''' | align="center" style="background:#f0f0f0;"|'''ng/ul''' | align="center" style="background:#f0f0f0;"|'''ng''' | align="center" style="background:#f0f0f0;"|'''ug''' | align="center" style="background:#f0f0f0;"|'''pmole''' | align="center" style="background:#f0f0f0;"|'''adaptor''' | align="center" style="background:#f0f0f0;"|'''ul (30uM)''' |- | PC2||0.91||16.4||0.016||0.12||2.42||0.08 |- | GFP+||0.29||5.27||0.005||0.04||0.76||0.03 |- | GFP-||0.23||4.14||0.004||0.03||0.6||0.02 |- | E9.5||0.4||7.22||0.007||0.05||1.06||0.04 |- | E11.5||1.61||28.98||0.029||0.22||4.4||0.15 |- | E13.5m||0.48||8.59||0.009||0.07||1.36||0.05 |- | E13.5f||2.72||48.96||0.049||0.37||7.42||0.25 |- | |} '''Purification with AMPure beads''' # 50ul beads to 50ul reaction, mix 10 times, R.T for 8min # Place on magnetic stand for 5min # Remove sup., wash with 180ul fresh made 80% EtOH twice # Air-dry for 15min, re-suspend with 22ul Qiagen EB, mix 10 times, RT 2min, magnetic stand 2min # Transfer 20ul DNA from beads, 1ul left in tube for Qubit measurement
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