Difference between revisions of "Team:TCU Taiwan/Result/vitro"

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<p>Aug. 6th ,2015</p>
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<p align="center"><font size="+3">signiferin</font></p>
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<p>Aug. 6th ,2015</p>
 
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&bull;&nbsp; * PCR signiferin
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&bull;&nbsp; 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.
 
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Revision as of 07:23, 18 September 2015


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

•  signiferin TA plasmid purification

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 plasmid.


Aug. 16th ,2015

•  pQE60 plasmid digest with enzyme BamH I and Nco I biobrick digest with enzyme EcoRI and Pst I

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

•  PCR Epi-1 insert
2% gel. It is the correct size that Epi-1 PCR product have band between 200~300 bp. But, there is also band on blank. We consider it was polluted when we add reagent.
•  plasmid purification of pQE60 vector with signiferin insert
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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