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MB Sample ID: SA023657
| Local Sample ID: | OG1RF_1 Decoy |
| Subject ID: | SU000489 |
| Subject Type: | Bacterial cells |
| Subject Species: | Enterococcus faecalis |
| Taxonomy ID: | 1351 |
| Genotype Strain: | OG1RF |
| Species Group: | Microorganism |
Select appropriate tab below to view additional metadata details:
Subject:
| Subject ID: | SU000489 |
| Subject Type: | Bacterial cells |
| Subject Species: | Enterococcus faecalis |
| Taxonomy ID: | 1351 |
| Genotype Strain: | OG1RF |
| Species Group: | Microorganism |
Factors:
| Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
|---|---|---|---|---|
| OG1RF_1 Decoy | SA023657 | FL005719 | Decoyinine, 0.1mg/mL | Exposure |
Collection:
| Collection ID: | CO000483 |
| Collection Summary: | After the 15 minute treatment interval, 20mL sample will be harvested by filtration onto 0.45uM nitrocellulose membrane. The cells will be scraped off of the membrane into ice-cold PBS (~1mL), centrifuged for 45 seconds at 12K RPM, liquid aspirated and stored at -80C until sending for analysis. |
| Sample Type: | Bacterial cells |
| Collection Method: | Fast filtration |
| Volumeoramount Collected: | 20ml |
| Storage Conditions: | -80C |
Treatment:
| Treatment ID: | TR000503 |
| Treatment Summary: | Triplicate cultures of E. faecalis OG1RF will be grown in FMC-AUG to OD600 = 0.25. The cultures will be split into 3 equal volumes of 30mL which will be exposed to 100uL DMSO, 0.01mg/mL mupirocin, or 0.1mg/mL decoyinine for 15 minutes. At this point, 10mL sample will be removed for protein analysis. The remaining 20mL will be harvested by fast filtration for analysis.|Triplicate cultures of E. faecalis OG1RF will be grown in FMC-AUG to OD600 = 0.25. The cultures will be split into 3 equal volumes of 30mL which will be exposed to 100uL DMSO, 0.01mg/mL mupirocin, or 0.1mg/mL decoyinine for 15 minutes. At this point, 10mL sample will be removed for protein analysis. The remaining 20mL will be harvested by fast filtration for analysis.|Triplicate cultures of E. faecalis OG1RF will be grown in FMC-AUG to OD600 = 0.25. The cultures will be split into 3 equal volumes of 30mL which will be exposed to 100uL DMSO, 0.01mg/mL mupirocin, or 0.1mg/mL decoyinine for 15 minutes. At this point, 10mL sample will be removed for protein analysis. The remaining 20mL will be harvested by fast filtration for analysis. |
| Treatment Compound: | DMSO|Mupirocin|Decoyinine |
| Treatment Dose: | 100uL|0.01mg/mL|0.1mg/mL |
| Treatment Doseduration: | 15 min|15 min|15 min |
| Treatment Vehicle: | DMSO|DMSO|DMSO |
Sample Preparation:
| Sampleprep ID: | SP000496 |
| Sampleprep Summary: | NAD-ATP |
| Sampleprep Protocol Filename: | SB_NAD-ATP_Assay_Agilent.pdf |
Combined analysis:
| Analysis ID | AN000731 |
|---|---|
| Analysis type | MS |
| Chromatography type | Reversed phase |
| Chromatography system | Agilent 1290 Infinity |
| Column | Thermo Hypercarb (50 x 2.1mm,3um) |
| MS Type | ESI |
| MS instrument type | Triple quadrupole |
| MS instrument name | Agilent 6490 QQQ |
| Ion Mode | POSITIVE |
| Units | nmol/mg protein |
Chromatography:
| Chromatography ID: | CH000524 |
| Chromatography Summary: | NAD-ATP |
| Methods Filename: | SB_NAD-ATP_Assay_Agilent.pdf |
| Instrument Name: | Agilent 1290 Infinity |
| Column Name: | Thermo Hypercarb (50 x 2.1mm,3um) |
| Chromatography Type: | Reversed phase |
MS:
| MS ID: | MS000648 |
| Analysis ID: | AN000731 |
| Instrument Name: | Agilent 6490 QQQ |
| Instrument Type: | Triple quadrupole |
| MS Type: | ESI |
| Ion Mode: | POSITIVE |
| Analysis Protocol File: | SB_NAD-ATP_Assay_Agilent.pdf |
