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MB Sample ID: SA113468
| Local Sample ID: | Evocrp4_5 |
| Subject ID: | SU001468 |
| Subject Type: | Bacteria |
| Subject Species: | Escherichia coli |
| Taxonomy ID: | 562 |
| Genotype Strain: | K-12 MG1655 |
Select appropriate tab below to view additional metadata details:
Subject:
| Subject ID: | SU001468 |
| Subject Type: | Bacteria |
| Subject Species: | Escherichia coli |
| Taxonomy ID: | 562 |
| Genotype Strain: | K-12 MG1655 |
Factors:
| Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
|---|---|---|---|---|
| Evocrp4_5 | SA113468 | FL014274 | Evolved | Genotype |
Collection:
| Collection ID: | CO001463 |
| Collection Summary: | The Wildtype, crp mutant and the five evolved replicate populations were grown in a bioreactor aerobically in glucose minimal media with 40 mM MOPS at 37 degrees Celsius and 700 rpm. The samples were harvested during its exponential phase of growth for three biological and two technical replicates (n=6). The metabolites were extracted using the Methanol-Chloroform-Water method. Detailed sample collection protocol is described in “extraction_method.pdf”. |
| Sample Type: | Bacterial cells |
| Collection Method: | Methanol-Chloroform-Water method |
| Collection Location: | IIT Bombay, Powai, Mumbai- 400076, Mumbai, Maharashtra |
| Storage Conditions: | -80℃ |
Treatment:
| Treatment ID: | TR001483 |
| Treatment Summary: | The Wildtype, crp mutant and the five evolved replicate populations were grown in a bioreactor aerobically in glucose minimal media with 40 mM MOPS at 37 degrees Celsius and 700 rpm. The metabolites were extracted at expoential phase to characterize and compare the metabolic profiles of all the strains. |
| Cell Growth Container: | Bioreactor |
| Cell Media: | M9 + Glucose + 40 mM MOPS |
| Cell Envir Cond: | 37 degrees Celsius, 700 rpm |
Sample Preparation:
| Sampleprep ID: | SP001476 |
| Sampleprep Summary: | Samples were extracted from the Wildtype, crp mutant and the five evolved replicate population, grown in a bioreactor aerobically in glucose minimal media with 40 mM MOPS at 37 degrees Celsius and 700 rpm. The metabolites were extracted using the Methanol-Chloroform-Water method. The samples were spiked with an equal volume of 13C-labelled internal standard taken from the same batch at an earlier stage to prevent to account for losses due to metabolite degradation and to enable robust quantification circumventing the ion-suppression effects. |
Combined analysis:
| Analysis ID | AN002332 | AN002333 |
|---|---|---|
| Analysis type | MS | MS |
| Chromatography type | HILIC | HILIC |
| Chromatography system | Thermo Dionex Ultimate 3000 | Thermo Dionex Ultimate 3000 |
| Column | SeQuant ZIC-HILIC (150 x 2.1mm,5um) | SeQuant ZIC-HILIC (150 x 2.1mm,5um) |
| MS Type | ESI | ESI |
| MS instrument type | Orbitrap | Orbitrap |
| MS instrument name | Thermo Q Exactive Plus Orbitrap | Thermo Q Exactive Plus Orbitrap |
| Ion Mode | POSITIVE | NEGATIVE |
| Units | Height ratio/gDCW | Height ratio/gDCW |
Chromatography:
| Chromatography ID: | CH001709 |
| Chromatography Summary: | The detailed chromatography and MS protocol has been described in "ms_method.pdf" file. |
| Instrument Name: | Thermo Dionex Ultimate 3000 |
| Column Name: | SeQuant ZIC-HILIC (150 x 2.1mm,5um) |
| Chromatography Type: | HILIC |
MS:
| MS ID: | MS002174 |
| Analysis ID: | AN002332 |
| Instrument Name: | Thermo Q Exactive Plus Orbitrap |
| Instrument Type: | Orbitrap |
| MS Type: | ESI |
| MS Comments: | Data acquisition done using parent ion MS intensities. Peak height used for processing. Xcalibur 4.3.73.11 used for data analysis and peak integration |
| Ion Mode: | POSITIVE |
| MS ID: | MS002175 |
| Analysis ID: | AN002333 |
| Instrument Name: | Thermo Q Exactive Plus Orbitrap |
| Instrument Type: | Orbitrap |
| MS Type: | ESI |
| MS Comments: | Data acquisition done using parent ion MS intensities. Peak height used for processing. Xcalibur 4.3.73.11 used for data analysis and peak integration |
| Ion Mode: | NEGATIVE |
