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MB Sample ID: SA163931
| Local Sample ID: | 33P |
| Subject ID: | SU001829 |
| Subject Type: | Bacteria |
| Subject Species: | Vibrio cholerae |
| Taxonomy ID: | 666 |
| Genotype Strain: | V. cholerae serotype O1 (strain ATCC 39315 / El Tor Inaba N16961- access:2018.06.26) |
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Subject:
| Subject ID: | SU001829 |
| Subject Type: | Bacteria |
| Subject Species: | Vibrio cholerae |
| Taxonomy ID: | 666 |
| Genotype Strain: | V. cholerae serotype O1 (strain ATCC 39315 / El Tor Inaba N16961- access:2018.06.26) |
Factors:
| Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
|---|---|---|---|---|
| 33P | SA163931 | FL019494 | OD0.5 | Condition |
| 33P | SA163931 | FL019494 | delta_VcdR_pVcdP | Strain |
Collection:
| Collection ID: | CO001822 |
| Collection Summary: | V. cholerae wild-type, ΔvcdRP and ΔgltA each harboring an empty control plasmid (pCtrl) and ΔvcdRP with pVcdRP expression plasmids (pVcdRP and pVcdP) as well as ΔgltA harboring pVcdP were grown in LB medium to exponential and stationary phase, and were subsequently quenched in ice-cold methanol. The quenched samples were centrifuged at 3000 x g for 10 min at -9°C. 1ml aliquot of supernatant was stored for leakage analysis, while the rest of the pellets were flash frozen in liquid nitrogen. |
| Sample Type: | Bacterial cells |
Treatment:
| Treatment ID: | TR001842 |
| Treatment Summary: | No treatment |
Sample Preparation:
| Sampleprep ID: | SP001835 |
| Sampleprep Summary: | The frozen cell pellets were thawed on ice and resuspended in 500 µl methanol. After three freeze (liquid nitrogen) and thaw cycles, the sample was centrifuged (16000 x g, -4 °C and 5 min). The supernatant was transferred into a new tube and stored on ice. The extraction step was repeated and the supernatants were combined prior to evaporation. The samples were stored at -80°C until further analysis. |
| Extract Storage: | -80℃ |
Combined analysis:
| Analysis ID | AN002855 |
|---|---|
| Analysis type | MS |
| Chromatography type | Ion pair |
| Chromatography system | Agilent 1290 Infinity II |
| Column | Waters XSelect HSS T3 XP (150 x 2.1mm,2.5um) |
| MS Type | ESI |
| MS instrument type | Triple quadrupole |
| MS instrument name | AB Sciex 6500+ QTRAP |
| Ion Mode | NEGATIVE |
| Units | relative abundance |
Chromatography:
| Chromatography ID: | CH002114 |
| Instrument Name: | Agilent 1290 Infinity II |
| Column Name: | Waters XSelect HSS T3 XP (150 x 2.1mm,2.5um) |
| Column Temperature: | 40 |
| Solvent A: | 5% methanol/2% isopropanol/93% water; 10 mM acetic acid; 10 mM tributylamine, pH 7.1 |
| Solvent B: | 100% isopropanol |
| Chromatography Type: | Ion pair |
MS:
| MS ID: | MS002648 |
| Analysis ID: | AN002855 |
| Instrument Name: | AB Sciex 6500+ QTRAP |
| Instrument Type: | Triple quadrupole |
| MS Type: | ESI |
| MS Comments: | MS acquisition as well as data processing software was Analyst vs 1.7.0 |
| Ion Mode: | NEGATIVE |
