Return to study ST000469 main page

MB Sample ID: SA023665

Local Sample ID:OG1RF_3E8
Subject ID:SU000490
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:SU000490
Subject Type:Bacterial cells
Subject Species:Enterococcus faecalis
Taxonomy ID:1351
Genotype Strain:OG1RF
Species Group:Microorganism

Factors:

Local Sample IDMB Sample IDFactor Level IDLevel ValueFactor Name
OG1RF_3E8SA023665FL005723WTStrain
OG1RF_3E8SA023665FL005723PBSResuspension

Collection:

Collection ID:CO000484
Collection Summary:3E9, 3E10 - upon reaching OD600 = 0.25, 100mL (OG1RF) or 425mL (rel/relQ) will be harvested by filtration onto 0.45uM nitrocellulose membrane. The cells will be scraped off of the membrane into ice-cold PBS or formic acid (~1 mL), centrifuged for 45 seconds at 12K RPM, liquid aspirated and stored at -80C until sending for analysis.|3E8 - (OG1RF only). Upon reaching OD600 = 0.25, 1mL culture will be harvested by 45 seconds centrifugation at 12K RPM, liquid decanted, and stored at -80C until sending for analysis.
Sample Type:Bacterial cells

Treatment:

Treatment ID:TR000504
Treatment Summary:This pilot study involves no treatment - cells will be grown in chemically defined medium to an OD600 of 0.25 and harvested for analysis

Sample Preparation:

Sampleprep ID:SP000497
Sampleprep Summary:NAD|NADH
Sampleprep Protocol Filename:SB_NAD-ATP_Assay_Agilent.pdf
SB_NADH_Assay_Agilent.pdf

Combined analysis:

Analysis ID AN000732 AN000733
Analysis type MS MS
Chromatography type Reversed phase Reversed phase
Chromatography system Agilent 1290 Infinity Agilent 1290 Infinity
Column Thermo Hypercarb (50 x 2.1mm,3um) Waters Acquity HSS T3 (100 x 2.1mm,1.8um)
MS Type ESI ESI
MS instrument type Triple quadrupole Triple quadrupole
MS instrument name Agilent 6490 QQQ Agilent 6490 QQQ
Ion Mode POSITIVE POSITIVE
Units nmol/mg protein nmol/mg protein

Chromatography:

Chromatography ID:CH000525
Chromatography Summary:NAD
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
  
Chromatography ID:CH000526
Chromatography Summary:NADH
Methods Filename:SB_NADH_Assay_Agilent.pdf
Instrument Name:Agilent 1290 Infinity
Column Name:Waters Acquity HSS T3 (100 x 2.1mm,1.8um)
Column Temperature:30
Flow Rate:0.54 mL/min
Solvent A:100% water; 5 mM ammonium acetate, pH 6
Solvent B:100% acetonitrile
Chromatography Type:Reversed phase

MS:

MS ID:MS000649
Analysis ID:AN000732
Instrument Name:Agilent 6490 QQQ
Instrument Type:Triple quadrupole
MS Type:ESI
MS Comments:NAD
Ion Mode:POSITIVE
  
MS ID:MS000650
Analysis ID:AN000733
Instrument Name:Agilent 6490 QQQ
Instrument Type:Triple quadrupole
MS Type:ESI
MS Comments:NADH
Ion Mode:POSITIVE
  logo