Part:BBa_J176171:Experience

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UNIQ552ba245bc35711c-partinfo-00000000-QINU UNIQ552ba245bc35711c-partinfo-00000001-QINU Hong Kong_HKUST iGEM 2013 planned to use this plasmid as backbone for transfecting in mammalian cells. However, we found that this plasmid is degradable at a high temperature during polymerase chain reaction.

Figure 1: Degradation of BBa_176171 plasmid at temperature above 50°C after denaturation step of polymerase chain reaction. BBa_176171 and pEGFP-N1 were placed in ddH2O and ran polymerase chain reaction denaturation step at 98°C for 30 seconds and placed in a range of temperature from 50°C to 59°C for 30minutes. For controls, BBa_176171 and pEGFP-N1 were stored in room temperature (24°C) and 4°C for 30minutes respectively. The plasmids were ran on Gel Red pre-stained 0.8% agarose gel using 140V for 25minutes. 0.5uL of GeneRuler 1kb ladder was used.