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Team:UNICAMP-EMSE Brazil/Notebook/1 September 2011
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Gel 1: flHDC+RBS+GFP C6a/ flHDC+RBS+GFP C6b/ flHDC+RBS+IL12 C8/ flHDC+IL 12 C10/ flHDC+IL 12 C11
1º September 2011
Checking digestion (E-P) of flHDC+RBS+IL12 and flHDC+RBS+GFP mini-preped today:
Objective:
- Checking digestion (E-P) of flHDC+RBS+IL12 and flHDC+RBS+GFP mini-preped today:
Samples:
- Gel 1:
- flHDC+RBS+GFP C6a/ flHDC+RBS+GFP C6b/ flHDC+RBS+IL12 C8/ flHDC+IL 12 C10/ flHDC+IL 12 C11
- Ladder:
- 100-10.000 bp (Fermentas)
- Gel Agarose concentration:
- 1,0%
- Results:
- Purification of SoxS+RBS+IL10 did not work. Just SoxS+RBS+HlyB
| Gene | Total size (gene+vector) | Linear vector size | Gene size | Result |
|---|---|---|---|---|
| flHDC+RBS+GFP C6a | 3661 bp | 2800 bp | 861 bp | Just vector |
| flHDC+RBS+GFP C6b | 3661 bp | 2800 bp | 861 bp | Just vector |
| flHDC+RBS+IL 12 C8 | 4584 bp | 2800 bp | 1784 bp | No bands |
| flHDC+RBS+IL 12 C10 | 4584 bp | 2800 bp | 1784 bp | No bands |
| flHDC+RBS+IL 12 C11 | 4584 bp | 2800 bp | 1784 bp | No bands |
- No colonies worked from plate with bacteria transformed with the ligation did on 27th. So we decided to repeat the ligation reaction (IL 12 or GFP with flhDC promoter) and transformation of this two genes. We changed the ligation approach for RBS+GFP, instead of ligating it with promoter we digested it with E-S and we are going to ligate with HlyA+T construct.
- Inoculation of the colonies in the plates done yesterday. Tranformation and platting of today´s ligation.
