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==Results== *3uL of amplified product was validated using TBE-PAGE **3uL WGA amplicon + 3uL 0.5X TBE + 3uL 6X loading dye. **Run at 200V, 30min on a 10-well TBE-PAGE (Use Invitrogen premade gel). **Long staing (15mins) with foil cover. **Picture was taken at exposure time 4.096sec [[File:sam050210-nextera lib.jpg|300px]] Concentration: 2h-Ecoli MDA(50ng) 10h-Ecoli MDA(50ng) QUBIT-HS (ng/uL) 17.0 15.4 *For using Low-molecular-weight(LMW) fragmentation buffer, the expeced size will be around 200-500bp. The result in gel picture support this point. *There still have some smear tail above 500bp. I am curious if it is caused by over PCR amplification. **Although the amplfication cycle (12 cycles) is suggested in protocol, I am inclined to test the result with 10cycles amplification using the remained 5uL templates. **A size selection may be necessary for removing large fragments. *NOTE: Condition summary **Starting material: 50ng MDA amplicon (non-sheared, but EtOH purified). **Using Zymo-DNA cleaning column **12cycle amplification (5uL template in 50uL rxn volume). *'''The similar observation (over-sized libraries) was found on Epicenter's website''' **http://epicentral.blogspot.com/search/label/nextera [[File:Nextera kit - oversize of libraries.jpg|500px]]
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