Metadata.txt

Version 1

# Metadata files
D:  
/SUSPHIRE_RMF  
/_p_SUSPHIRE  
/_I_T21_SxPv1Analysis  
/_S_SxPv10vsSxP12  
/_A_RootsSxPv10vsv12-GCMS

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* **/_ASSAY_METADATA.TXT**
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Item	Value 
Assay:	_A_RootsSxPv10vsv12-GCMS 
Short Name:	RootsSxPv10vsv12-GCMS 
Assay Class:	WET 
Assay Type:	GCMS 
Title:	Roots volatilome of SxPv1.0, SxPv1.2 T1 and WT 
Description:	he purpose of this assay is to define the volatilome of SxPv1.0, v1.2 and WT Nicotiana benthamiana plant roots, focusing on the differences between them, by GC-MS. 
pISA Assay creation date:	2021-11-25 
pISA Assay creator:	RMF 
Lab manager:	DO 
Sample collection protocol:	Root samples were collected at 7 weeks, in a early flowering stage of plants. Samples were collected between 4 and 6 pm, frozen in liquid nitrogen immediately after collection, and ground afterwards. 
Extraction protocol:	50 mg of frozen, ground root samples were weighed in a 10mL headspace screw-cap vial and stabilized by adding 1mL of 5M CaCl2 and 150 æL of 500mM EDTA (pH = 7.5), after which they were sonicated for 5 minutes. Volatile compounds were captured by means of headspace solid phase microextraction (HS-SPME) with a 65 æm polydimethylsiloxane/divinylbenzene (PDMS/DVB) SPME fiber (Supelco, Bellefonte, PA, USA). Volatile extraction was performed automatically by means of a CombiPAL autosampler (CTC Analytics). Vials were first incubated at 80øC for 3 minutes with 500 rpm agitation. The fiber was then exposed to the headspace of the vial for 20 min under the same conditions of temperature and agitation. Desorption was performed at 250øC for 1 minute (splitless mode) in the injection port of a 6890N gas chromatograph coupled to a 5975B mass spectrometer (Agilent Technologies). After desorption, the fiber was cleaned in a SPME fiber conditioning station (CTC Analytics) at 250øC for 5 min under a helium flow. 
Chromatography protocol:	Chromatography was performed on a DB5ms (60 m, 0.25 mm, 1 µm) capillary column (JandW) with helium as the carrier gas at a constant flow of 1.2mLxmin-1. The oven conditions started with an initial temperature of 160°C for 2 min, 7°Cmin-1 ramp until 280°C, and a final hold at 280°C for 6 minutes. 
Mass spectrometry protocol:	Electron impact ionization (EI), 70 eV ionization energy, MS source temperature 230°C, MS quadrupole temperature 150°C, single quadrupole detector, m/z range 35-300. 
Phenodata:	../../phenodata_20210115.txt 
Featuredata:	 
Creation date:	2021-01-15 
Extract ID:	$_extr 
Extraction Method:	HS-SPME 
Date Extraction:	2021-01-15 
Derivatization or Labelling:	none 
Date Derivatization or Labelling:	2021-01-15 
Derivatized or labeled Extract ID:	$_extrD 
Other Post Extraction Procedures:	 
Storage:	 
Date GC-MS Run:	2021-01-15 
GC Instrument:	6890N gas chromatograph (Agilent Technologies) 
GC Autosampler Model:	CombiPAL autosampler (CTC Analytics) 
GC Column model:	DB5ms (60m, 0.25 mm, 1um) capillary column (JandW) 
GC Column type:	capillary column 
Guard Column:	 
MS Scan polarity:	positive 
MS Scan mz range:	35-300 
MS Instrument:	5975B mass spectrometer (Agilent Technologies) 
MS Ion source:	electron ionization (EI) 
Mass analyzer:	quadrupole mass filter 
Operator:	RMF 
Notes:	 
Data:	 

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