pSB1C3 plasmid gel electrophoresis

Aims

The aim of this experiment is to run gel electrophoresis for the extracted and linearized plasmid pSB1C3 fragment which were amplified at 4 different melting temperatures by 4 separate PCR reactions.

Materials and Protocol

Please refer to: gel electrophoresis for gel electrophoresis protocol.

Result

Figure 1: Gel electrophoresis of the amplified linearized plasmid pSB1C3 fragments ran at 4 different melting temperatures, Tms, (50, 60, 65, 70°C). A 1 kb DNA ladder was used on either side of lanes.

• Lane 1: pSB1C3 fragment amplified at 55°C
• Lane 2: pSB1C3 fragment amplified at 60°C
• Lane 3: pSB1C3 fragment amplified at 65°C
• Lane 4: pSB1C3 fragment amplified at 70°C
• Lane 5: pSB1C3 fragment amplified at 55°C
• Lane 6: pSB1C3 fragment amplified at 60°C
• Lane 7: pSB1C3 fragment amplified at 65°C
• Lane 8: pSB1C3 fragment amplified at 70°C

Discussion

No bands were found in any of the lanes. Yesterday, a faint band was found when the melting temperature was set at 65°C but today no band is found in lane 3 and lane 7. This makes finding the cause for no amplification even difficult. We would still be looking into it and would be changing other parameters.

Conclusion