Difference between revisions of "Team:Hong Kong UCCKE/Results"

After we have successfully cloned our plasmids into the E.coli, we did miniprep to obtain purified plasmid DNA. Then, we did restriction map and have sent the samples to BGI for sequencing. The results are shown below.

BBa_K2197300

Restriction Map

After mini prep, we use the same restriction enzyme EcoR1 and Pst1 to do restriction digestion and gel electrophoresis. However, the size of K2197300 and the backbone pSB1C3 is almost the same, thus we cannot prove by the Gel photo.

Image of gel electrophoresis of 300a+b

Sequencing Result

With the help of CUHK, we sent our plasmid to BGI for sequencing. We aligned the sequence that we ordered from IDT(As the template sequence 99593011 below) with the sequencing result received from BGI(as the aligned sequence below). And here is our analysis.

The black area indicates that the two sequence are the same which those red strip represent that there're some mutations.

Sequencing result of BBa_K2197300

Sequence Alignment(1)

Sequence Alignment(2)

Conclusion

From the results above, we can say we successfully clone the DNA that IDT sent us into the cell. However, as what we have mentioned in the experiment page, we found that the cells cannot express GFP. Then, we aligned the sequence registered in IGEM with the one that IDT send us, we found that the sequence containing 'tactagag' is missing.

BBa_K2197400

Restriction Map

After mini prep, we use the same restriction enzyme EcoR1 and Pst1 to do restriction digestion and gel electrophoresis. As seen, the size of K2197400 (2214 bp) and the backbone pSB1C3 (2070 bp)is almost the same, similar to our predicted result.

Image of gel electrophoresis of 400a+b and 500

Image of theoretical gel electrophoresis of 400 from genome compiler

Sequencing Result

Sequencing Result of BBa_K2197400

Sequence Alignment (1)

Sequence Alignment (2)

Conclusion

From the results above, we can say we successfully clone the plasmid DNA into the cell.

BBa_K2197500

Restriction Map

Also using the same restriction enzyme EcoR1 and Pst1 to do restriction digestion and gel electrophoresis. The size of K2197500 (1668 bp) and the backbone pSB1C3 (2070 bp).

Image of gel electrophoresis of 400a+b and 500

Image of theoretical gel electrophoresis of 500 from genome compiler

Project 500 (1)

Project 500 (2)