Team:NCKU Tainan/Notebook Construction
(pSB1C3-BBa_B0034, pSB1C3-BBa_B0015, pSB1C3-BBa_K592009)
2016.03.30
We got the bacteria with Biobrick from XMU.
2016.04.01
Pre-culture: 3 ml of LB (with Chloramphenicol) was inoculated with one colony from LB agar plate.
2016.04.02
Culture: 10 ml of LB (with Chloramphenicol) culture with 100 ml/μL pre-cultured bacteria.
And grow up at 37 degree for 10-12 hours.
2016.04.03
Extracted the plasmid, and measured the concentration.
Store the plasmid at -20 degree.
2016.04.16
PCR for PpI & Pcar.
Result: After gel purified, the concentration of the DNA was too low to do construction.
2016.04.17
PCR for PpI & Pcar again (in big scale).
Purified the PCR product and cloned to T-vector.
2016.04.18
Confirmed the white colony by PCR (using M13F & M13R as the primer)
Result: Some of the colonies were correct.
Pre-culture: 3 ml of LB (with Ampicillin) was inoculated with one correct colony from the agar plate.
2016.04.19
Extracted plasmid and confirmed plasmid size by gel electrophoresis.
Result: The size of pMD19T-PpI and pMD19T-Pcar were correct.
2016.04.20
Culture: 10 ml of LB (with Ampicillin) cultured with 100 ml/μL pre-cultured bacteria.
Grow up at 37 degree for 8 hours and stored the bacteria at -70 refrigerator.
2016.04.30
Digested pSB1C3-B0034 (SpeI,PstI) and pSB1C3-K592009 (XbaI,PstI)
Ligated K592009 (insert) to pSB1C3-B0034 (vector) and transformed the product.
2016.05.01
Confirmed by colony PCR (using Prefix-F & Suffix-R as the primer).
Pre-culture: 3 ml of LB (with Chloramphenicol) was inoculated with one correct colony from the agar plate.
2016.05.02
Extracted plasmid and confirmed plasmid size by gel electrophoresis.
Result: The size of pSB1C3-B0034-K592009 was correct.
2016.05.03
Culture: 10 ml of LB (with Chloramphenicol) cultured with 100ml/μL pre-cultured bacteria.
Grow up at 37 degree for 8 hours and stored the bacteria at -70 refrigerator.
2016.05.07
Digested pSB1C3-B0034-K592009 (XbaI,PstI), pMD19T-Pcar (SpeI,PstI) and pMD19T-PpI (SpeI,PstI)
Ligated B0034-K592009 (insert) to pMD19T-Pcar (vector) and pMD19T-Pcar (vector).
Transformed the product.
2016.05.08
Confirmed by colony PCR (using M13F & M13R as the primer).
Pre-culture: 3 ml of LB (with Ampicillin) was inoculated with one correct colony from the agar plate.
2016.05.09
Extracted plasmid and confirm plasmid size by gel electrophoresis.
Result: The size of pMD19T-Pcar-B0034-K592009 and pMD19T-PpI-B0034-K592009 were correct.
2016.05.10
Culture: 10 ml of LB (with Ampicillin) cultured with 100 ml/μL pre-cultured bacteria.
Grow up at 37 degree for 8 hours and stored the bacteria at -70 refrigerator.
(BBa_J23110, BBa_J23101,BBa_B0031,BBa_B0032,BBa_E1010)
2016.06.03
Transformed 3 ml/μL biobrick to 100 ml/μL DH5α competent cell by heat shock.
Result: The transformation of BBa_J23110, BBa_B0032 and BBa_E1010 was successful.
Result: The transformation of BBa_J23110, BBa_B0032 and BBa_E1010 was fail.
2016.06.04
Pre-culture: 4 ml of LB (with Chloramphenicol) was inoculated with one colony from LB agar plate.
Extracted plasmid by Plasmid miniPREP Kit and confirmed plasmid size by gel electrophoresis.
2016.06.05
Culture: 10 ml of LB (with Chloramphenicol) was inoculated with one colony from LB agar plate. And grow up at 37 degree for 10-12 hours.
2016.06.06
Extracted the plasmid, and measure the concentration.
Stored the plasmid at -20 degree.
2016.06.07
Transformed 5 ml/μL biobrick to 100 ml/μL DH5α competent cell by heat shock.
(BBa_J23110, BBa_B0032 and BBa_E1010)
Result: The transformation of BBa_J23110, BBa_B0032 and BBa_E1010 were successful.
2016.06.08
Pre-culture: 4 ml of LB (with Chloramphenicol) was inoculated with one colony from LB agar plate.
Extracted plasmid by Plasmid miniPREP Kit and confirmed plasmid size by gel electrophoresis.
2016.06.09
Culture: 10 ml of LB (with Chloramphenicol) was inoculated with one colony from LB agar plate.
And grow up at 37 degree for 10-12 hours.
2016.06.10
Extracted the plasmid, and measure the concentration.
Stored the plasmid at -20 degree.
