Team:TCU Taiwan/Result/vitro

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signiferin

Aug. 6th ,2015

•  We did PCR of signiferin. After doing gel electrophoresis to make sure the size, we did gel extraction, ligated the PCR product into TA clone, transformed it into competent cell, and culture it.

2% gel. The PCR product of signiferin should be 100~200 bp. As the result, there is band between 100~200 bp on sample 2 so we consider it is correct.

Aug. 15th ,2015

• We purified the plasmid of TA clone with signiferin, and did gel electrophoresis to make sure the size.

0.8% gel. The size of TA plasmid with signiferin insert should be 2000~3000 bp. In addition, plasmid will become sorpercoil so it will run faster. Therefore, we consider these are the correct plasmids.

Aug. 16th ,2015

• We used BamHI and NcoI these two kinds of enzyme to check the insert really contain in TA clone. After doing gel electrophoresis to make sure the size, we did gel extraction, ligated it into backbone, transformed into competent cell, and culture it.

2% gel. The size of signiferin insert should be 100~20 bp. In addition, the signiferin insert in biobrick should be a little bigger than in pQE60. As the result, these two are all the correct sample we need.

Aug. 21st ,2015

• We pureified the plasmid of pQE60 with signiferin, and did gel electrophoresis to make sure the size.

0.8% gel. The size of our plasmid should be 3000 to 4000. As the result, these samples are the plasmid we need.

Aug. 22rd ,2015

•  Plasmid purification of pQE60 plasmid with signiferin insert •  Plasmid purification of backbone with signiferin insert

0.8％ gel. The size of signiferin biobrick should be 2000~3000 bp, and the size of pQE60 plasmid with signiferin should be 3000~4000 bp. In addition, the plasmid will become surpercoil so it will run faster. Therefore, they are all correct pasmid.

Aug. 23rd ,2015

•  Enzyme digestion to check the clone of biobrick with signiferin

2％ gel. The size of signiferin insert should be 100 to 200 bp. There is band on the sample 2 of enzyme digest product. As the result, we can make sure that the signiferin has been ligated in backbone.

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