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Team:Evry/Notebook/Sensing/PCBs/08-20-2014
From 2014.igem.org
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<u>Survival test on E.coli BL21:</u> | <u>Survival test on E.coli BL21:</u> | ||
| - | <br/>Bacteria survive again for different concentrations but they were very concentrated | + | <br/>Bacteria survive again for different concentrations but they were very concentrated |
| - | <br/> | + | <br/> |
| - | <br/> A dilution of the medium has been done for the positive control and the six different concentrations: | + | <br/> A dilution of the medium has been done for the positive control and the six different concentrations:<br/><img src="https://static.igem.org/mediawiki/2014/3/3a/2nd_Survival_test_1st_day.jpg" alt="image not found" /> |
</p> | </p> | ||
Revision as of 13:08, 8 October 2014
Sensor construction bphR2/PbphR1
We received sequencing reads: that match perfectly to the registry sequence.
26DJ54
AATAGGCGTTATCACGAGGCAGAANTTCAGATAAAAAAAATCCTTAGCTTTCGCTAAGGATGATTTCTGGAATTCGCGGCCGCTTCTAGAGATGTCCCTGG
GTGACATGCGTGATTTGGCCGCCACGCGGATCGCGCTCGAGAGCGAAGCGTTACGCCAAAGCGTGCTGAATGGTGACGCTGAATGGGAGGCGCGGATCGTCAGTTC
GTTTCACCGACTGTCATTGATTGAAGAGCCCACGATGCGGGATCCGGCTCGCTGGTTTAATGAGTGGGAGCCAGTCAACCGCGGTTTTCACGAAGCTCTTATCTCT
GCCTGTTCGTCCGTCTGGATCCGGCGGTTCCTGTCCATCCTGTATGTGCATATGGAGCGCTACCGCCGATTGACTGCTTACTAGTAGCGGCCGCTGCAGTCCGGCA
AAAAAGGGCAAGGTGTCACCACCCTGCCCTTTTTCTTTAAAACCGAAAAGATTACTTCGCGTTATGCAGGCTTCCTCGCTCACTGACTCGCTGCGCTCGGTCGTTC
GGCTGCGGCGAGCGGTATCA
GCTCACTCAGGG
26DJ55
CGAGTCAGTGAGCGAGGAAGCCTGCATAACGCGAAGTAATCTTTTCGGTTTTAAAGAAAAAGGGCAGGGTGGTGACACCTTGCCCTTTTTTGCCGGACTGC
AGCGGCCGCTACTAGTAAGCAGTCAATCGGCGGTAGCGCTCCATATGCACATACAGGATGGACAGGAACCGCCGGATCCAGACGGACGAACAGGCAGAGATAAGAG
CTTCGTGAAAACCGCGGTTGACTGGCTCCCACTCATTAAACCAGCGAGCCGGATCCCGCATCGTGGGCTCTTCAATCAATGACAGTCGGTGAAACGAACTGACGAT
CCGCGCCTCCCATTCAGCGTCACCATTCAGCACGCTTTGGCGTAACGCTTCGCTCTCGAGCGCGATCCGCGTGGCGGCCAAATCACGCATGTCACCCAGGGACATC
TCTAGAAGCGGCCGCGAATTCCAGAAATCATCCTTAGCGAAAGCTAAGGATTTTTTTTATCTGAAATTCTGCCTCGTGATACGCCTATTTTTATAGGTTAATGTCA
TGATAATAATGGTTTCTTAGA
Plasmid with bphR2 gene was purified with NucleoSpin Plamsid protocol (Macherey-Nagel) => we obtained 227,5ng/µL
This plasmid was digested according this protocol:
- Add:
- sterilized water: qsp 20µL
- template DNA : 500ng
- buffer 2.1: 2µL
- BSA: 0,2µL
- EcoRI: 1µL
- PstI: 1µL
- Reverse for mix
- Incubate at 37°C during 45mn
- Incubate at 80°C during 20mn
- Ligation was not possible because pSB1C3 plasmid was not digested so the digested plasmid (bphR2 gene) was stored at -20°C
Survival test on E.coli BL21:
Bacteria survive again for different concentrations but they were very concentrated
A dilution of the medium has been done for the positive control and the six different concentrations:
Aug 20
