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==Results== *3uL of amplified product was validated using TBE-PAGE *3uL amplified libraries were validated using 6%TBE-PAGE with standard long staining method(250V, 20min). **3uL WGA amplicon + 3uL 0.5X TBE + 3uL 6X loading dye. [[File:sam080510- Nextera lib amp - phusion EZ.jpg|600px]] [[File:sam080610- Nextera-lib amplified.jpg|300px]] *The templates from these qPCR amplification is 10ng **Start with 20ng but only 1/2 of 1-step product were amplified. **The sample 10h-4 may contain more template than 90m-4 so it was amplified faster. **The sample 10h-4 was prepared by dilution method to get estimated 20ng as starting amount. It is possible that the inaccuracy of quantification cause higher template used in 10h-4 sample. *Using 10ng templates for qPCR is not stable in this method, even with 16cycle amplification the PCR amplicons still hard to be seen. However if I keep amplifying the background may start showing up.
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