Sequencing analysis determines the order in which base pairs are arranged. In this project, we gave our samples to the company that sent them for sequencing, with the aim of using sequencing technology to look at statistical mutations and thus test whether our Generation II black box system is functioning properly. We first designed experiments to transform the first and second generation plasmids into BL21 E. coli for mutation respectively and set up a light gradient, we then sent the bacteria from these experiments for sequencing to verify the operation of the black box system. We then mutated key enzymes in the pentose five-carbon phosphate pathway (PPP), the tricarboxylic acid cycle (TCA), and the spermidine synthesis pathway in E. coli with the first and second generation plasmids, and sent the samples for sequencing as well, to observe the results of the mutations. Below are our sequencing results for these samples.
1.First generation-plasmid sequence analysis
2.Second generation-plasmid sequence analysis
3.Spermidine-plasmid sequence analysis
4.Nicotinate-plasmid sequence analysis