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==Discussion== *Nine out of 24 reactions (with 0.5 cell/uL template) were pick up for regular PCR confirmation ** A3, A6, A7, B4, B7, C3, C4 and C8 *One Blank reaction (D8) failed due to insufficient Master mix. Need to increase the master mix volume for this design. *The back ground amplification (Ct=26~28)still occur in Blank control, althoough the UV-treated H2O (from phi29 kit) was used as template. **ALS and NS buffer were UV-treated. All of the phi29 reagents kit except dNTP and enzyme were UV-treated (10 min). **It is possible the contamination is from the Master mix.
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