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MB Sample ID: SA031601

Local Sample ID:nC18-13nov15-027-r002
Subject ID:SU000607
Subject Type:Zebrafish
Subject Species:Danio rerio
Taxonomy ID:7955
Species Group:Fish

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Subject:

Subject ID:SU000607
Subject Type:Zebrafish
Subject Species:Danio rerio
Taxonomy ID:7955
Species Group:Fish

Factors:

Local Sample IDMB Sample IDFactor Level IDLevel ValueFactor Name
nC18-13nov15-027-r002SA031601FL007187malesex
nC18-13nov15-027-r002SA031601FL007187muscletissue
nC18-13nov15-027-r002SA031601FL00718748h post feedingtimepoint
nC18-13nov15-027-r002SA031601FL007187WTgenotype

Collection:

Collection ID:CO000601
Collection Summary:When testing the wildtype fasting fish, we only choose male fish. For each of the 3 time points, we scarifice 25 fish, put 5 tissue in one tube for 5 repeats. Because of tissue size, I adjusted heart sample at 2h and kidney sample at 2h to 3 repeats, liver sample at 2h to 4 repeats. For tfeb-/-fish, we collected heart, muscle, kidney with each of them 5 repeats. We collected both male and female. For lmna-/- we collected heart and muscle(only male) with each 3 repeats.
Sample Type:Blood

Treatment:

Treatment ID:TR000621
Treatment Summary:This is pre-test study for protocal development and baseline measures. Fish hearts are small, and more than 30 adult fish heart might need to be pooled to generated sufficient sample for a metabolic profiling experiments. Therefore, we will first test the condition using wild type fish at 3 months for this purpose. Tfeb -/- knockouts and lmna -/- knockouts are similarily being used for protocal development. After we optimize the protocol, we will assess changes in DOX-induced cardiomyopathy.

Sample Preparation:

Sampleprep ID:SP000614
Sampleprep Summary:Handling of tissue 1) Keep tubes on ice for 10 min. Spin down @ 1000 RCF for 30 sec 2) Add 20 ul PBS per sample and 3 ul 13C6-Phe 3) Sonicate on ice bath for 30 sec x 4 times 4) Add 250 ul ACN/MeOH mixture and incubate on ice for 30 min 5) Pool 20 ul sup from each sample and split 100 ul pool in 5 tubes 6) Split remaining sup in two 1 dram vials x 2 (100 ul per vial) 7) Dry under N2 stream

Combined analysis:

Analysis ID AN000896 AN000897 AN000898 AN000899
Analysis type MS MS MS MS
Chromatography type HILIC HILIC Reversed phase Reversed phase
Chromatography system Agilent 1290 Infinity Agilent 1290 Infinity Agilent 1290 Infinity Agilent 1290 Infinity
Column Waters Acquity BEH Amide (150 x 2.1mm,1.7um) Waters Acquity BEH Amide (150 x 2.1mm,1.7um) Waters Acquity HSS C18 (150 x 2.1mm,1.8um) Waters Acquity HSS C18 (150 x 2.1mm,1.8um)
MS Type ESI ESI ESI ESI
MS instrument type QTOF QTOF QTOF QTOF
MS instrument name Agilent 6550 QTOF Agilent 6550 QTOF Agilent 6550 QTOF Agilent 6550 QTOF
Ion Mode POSITIVE NEGATIVE POSITIVE NEGATIVE
Units unspecified unspecified unspecified unspecified

Chromatography:

Chromatography ID:CH000638
Instrument Name:Agilent 1290 Infinity
Column Name:Waters Acquity BEH Amide (150 x 2.1mm,1.7um)
Chromatography Type:HILIC
  
Chromatography ID:CH000639
Instrument Name:Agilent 1290 Infinity
Column Name:Waters Acquity HSS C18 (150 x 2.1mm,1.8um)
Chromatography Type:Reversed phase

MS:

MS ID:MS000798
Analysis ID:AN000896
Instrument Name:Agilent 6550 QTOF
Instrument Type:QTOF
MS Type:ESI
Ion Mode:POSITIVE
  
MS ID:MS000799
Analysis ID:AN000897
Instrument Name:Agilent 6550 QTOF
Instrument Type:QTOF
MS Type:ESI
Ion Mode:NEGATIVE
  
MS ID:MS000800
Analysis ID:AN000898
Instrument Name:Agilent 6550 QTOF
Instrument Type:QTOF
MS Type:ESI
Ion Mode:POSITIVE
  
MS ID:MS000801
Analysis ID:AN000899
Instrument Name:Agilent 6550 QTOF
Instrument Type:QTOF
MS Type:ESI
Ion Mode:NEGATIVE
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