Team:Ryerson Toronto/Results
Results
As shown by the agarose gel, instead of a clean band around 6000kbs as expected we observed multiple bands that were below 500kbs.
Looking Forward...
A general limitation of PCR is its inability to amplify fragments larger than 3kb. We are hoping to use two troubleshooting techniques to overcome this barrier. The first is to adopt the PCR protocol utilized by Ponce et al. to amplify DNA fragments of 6kp (1992). If this was unsuccessful, the second would be to design primers for three groups of genes and amplify them separately. These genes will be ligated in to a single plasmid and will be transformed in to the Escherichia coli strain DH5α as a primary propagation host. Afterwards, using a plasmid mini preparation kit, the plasmids would be purified. The purified plasmids would be transformed in to the cyanobacteria strain PCC 6803.
Ponce MR, Micol JL. PCR amplification of long DNA fragments. Nucleic Acids Research. 1992;20(3):623.
