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Team:NTNU Trondheim/Notebook
From 2014.igem.org
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Team:Cornell/notebook
From 2013.igem.org
Notebook
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Week 23
(02/06 - 08/06)
June 3rd
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testtesttesttest test
- Prepared SOC and yB solutions for future lab work.
- Sterilized material and solutions needed for future lab work.
June 4th
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Made LB plates with ampicillin and ampicillin + kanamycin.
June 5th
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Inoculated E. coli DH5α in SOC medium overnight.
June 6th
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OD of culture was 0.3160 after 100 minutes.
Made competent E. coli DH5α cells.
Week 24
(09/06 - 15/06)
June 10th
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Transformation was done by heat-shock. A plasmid containing ampicillin resistance was used, and the transformed cells were incubated overnight on LB plates with ampicillin. Plates showed a bacterial blanket the next day; the cells were apparently super competent.
Test of transformation efficiency of competent E. coli DH5α from June 6th.
June 11th
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Only 1 µl was used for the transformation, the rest of the BioBrick stock solution was stored -20 °C. BBa_J23101 was located at plate 4, 17F. BBa_B0034 was located at plate 4, 1N and BBa_C0012 was located at plate 4, 1P. Plates showed decent growth the next day; transformation a success.
Rehydrated BioBrick BBa_J23101, BBa_B0034 and BBa_C0012, and transformed them into competent E. coli DH5α cells by heat-shock, and incubated the cultures on LB plates with ampicillin overnight.
June 12th
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The SOC medium was clear the next day, meaning no growth.
Inoculated BioBrick colonies from June 11th in SOC medium containing ampicillin.
June 13th
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The failed inoculation attempt on June 12th could indicate something wrong with the LB plates with ampicillin. Growth of non-transformed cells supported this. Most likely the ampicillin was aliquotted to the medium at an elevated temperature, causing denaturation of the antibiotic.
Negative control of non-transformed E. coli DH5α on LB plates with ampicillin.
