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MB Sample ID: SA113117
| Local Sample ID: | POS_12 |
| Subject ID: | SU001461 |
| Subject Type: | Mammal |
| Subject Species: | Myotis lucifugus |
| Taxonomy ID: | 59463 |
Select appropriate tab below to view additional metadata details:
Subject:
| Subject ID: | SU001461 |
| Subject Type: | Mammal |
| Subject Species: | Myotis lucifugus |
| Taxonomy ID: | 59463 |
Factors:
| Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
|---|---|---|---|---|
| POS_12 | SA113117 | FL014238 | Pd | Treatment |
Collection:
| Collection ID: | CO001456 |
| Collection Summary: | Laboratory experiments were conducted at the University of Winnipeg (Manitoba Sustainable Development Species at Risk/Wildlife Scientific Permit # SAR16009). M. lucifugus were collected from a WNS-negative hibernaculum in Central Manitoba in January (mid-winter) 2017. E. fuscus were collected from a WNS-negative hibernaculum in northwestern Ontario near Kenora in the same month. Hibernating bats were removed from the cave wall by hand and transferred to a bio-secure bat facility as previously described (Warnecke et al., 2012; McGuire et al., 2016). Bats were tested to be negative for Pd upon arrival using standard qPCR techniques and inoculated using previously established protocols (Warnecke et al., 2012; McGuire et al., 2016). |
| Sample Type: | Liver |
Treatment:
| Treatment ID: | TR001476 |
| Treatment Summary: | Bats given sham (PBST) or Pd inoculation |
Sample Preparation:
| Sampleprep ID: | SP001469 |
| Sampleprep Summary: | Liver samples (~10 mg) were homogenized with cold methanol (300 μl) containing internal standards (10 μl SPLASH® Lipidomix®), 10 μl was removed for protein quantification, and the tissue homogenate was incubated on ice for 5 min. Chilled chloroform (600 μl) was added to samples and followed by shaking (vortex 30 sec) and further incubation on ice (10 min). Chilled water (300 μl) was added to samples followed by shaking (vortex 30 sec) and incubation on ice (10 min), then centrifuged for 10 min (10,000 x g, 4°C) to separate layers. The lower organic phase was separated, evaporated under vacuum, and reconstituted in 200 μl isopropanol (IPA):acetonitrile (ACN):H2O (2:1:1) for analysis. |
| Processing Storage Conditions: | On ice |
| Extract Storage: | On ice |
Combined analysis:
| Analysis ID | AN002315 | AN002316 |
|---|---|---|
| Analysis type | MS | MS |
| Chromatography type | Reversed phase | Reversed phase |
| Chromatography system | Waters Acquity | Waters Acquity |
| Column | Waters Acquity CSH C18 (100 x 2.1mm,1.7um) | Waters Acquity CSH C18 (100 x 2.1mm,1.7um) |
| MS Type | ESI | ESI |
| MS instrument type | QTOF | QTOF |
| MS instrument name | Waters Synapt G2 QTOF | Waters Synapt G2 S QTOF |
| Ion Mode | POSITIVE | NEGATIVE |
| Units | m/z values | m/z values |
Chromatography:
| Chromatography ID: | CH001702 |
| Methods Filename: | epannkuk_chromatography.docx |
| Instrument Name: | Waters Acquity |
| Column Name: | Waters Acquity CSH C18 (100 x 2.1mm,1.7um) |
| Chromatography Type: | Reversed phase |
MS:
| MS ID: | MS002158 |
| Analysis ID: | AN002315 |
| Instrument Name: | Waters Synapt G2 QTOF |
| Instrument Type: | QTOF |
| MS Type: | ESI |
| MS Comments: | None |
| Ion Mode: | POSITIVE |
| MS ID: | MS002159 |
| Analysis ID: | AN002316 |
| Instrument Name: | Waters Synapt G2 S QTOF |
| Instrument Type: | QTOF |
| MS Type: | ESI |
| MS Comments: | None |
| Ion Mode: | NEGATIVE |
| Analysis Protocol File: | epannkuk_processing.docx |
