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===Tagmentation reaction=== *Set up tagmentation reaction on cold block: 1rxn 4.5rxn DNA template+H2O 15.0 - 5X Nextera Rxn buf.(LMW) 4.0 18.0 Nextera enzyme 1.0 4.5 ------------------------------------- 20.0 22.5/4.5 = 5uL -- template 15uL *Briefly vortexing and centrifuging. Incubate in thermocycler (using instant incubation) at 55C for 5min. *During the incubation, setup the tube and buffer of MiniElute columns. *Reaction products (20uL) were purified using MiniElute column following the standard QIAquick protocol. **MiniElute columns have lids which are better to avoid introduced contamination. **The efficiency of MiniElute columns has been verified in Nextera-kit manual. *Elute DNA in 11uL pre-warmed '''EB buffer'''.
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