_ASSAY_METADATA.TXT

Version 1

Assay:	_A_TransientSPv11andSPv12-GCMS
Short Name:	TransientSPv11andSPv12-GCMS
Assay Class:	WET
Assay Type:	GCMS
Title:	Transient expression of constructs for SxPv1.1 and SxPv1.2
Description:	The constructs for SxPv1.1 and SxPv1.2 were tested transiently in Nicotiana benthamiana leaves. Pheromone content was analysed via GC-MS
pISA Assay creation date:	2020-07-23
pISA Assay creator:	ElenaMG
Lab manager:	DO
Sample collection protocol:	Leaf samples were collected at 4 weeks after transplant for young stage and at 7 weeks for adult stage (early flowering). Samples were collected between 4 and 6 pm, frozen in liquid nitrogen immediately after collection, and ground afterwards.
Extraction protocol::	50mg of frozen, ground leaf samples were weighed in a 10mL headspace screw-cap vial and stabilized by adding 1mL of 5M CaCl2 and 150 μL 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 μm polydimethylsiloxane
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. For an initial identification of the pheromone peaks, oven programming conditions were 40°C for 2 min, 5°Cmin-1 ramp until reaching 280°C, and a final hold at 280°C for 5 min. Once the target peaks were identified, the oven conditions were changed to 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 to reduce the overall running time without losing resolution of the desired compounds 
Mass spectrometry protocol::	Electron impact ionization (EI), 70 eV ionization energy, MS source temperature 230ºC, MS quadrupole temperature 150ºC, single quadrupole detector, m
Phenodata:	../../phenodata_20150819.txt
Featuredata:	
Creation date:	2020-07-23
Extract ID:	$_extr
Extraction Method:	HS-SPME
Date Extraction:	2020-07-22
Derivatization or Labelling:	none
Date Derivatization or Labelling:	2020-07-22
Derivatized or labeled Extract ID:	$_extrD
Other Post Extraction Procedures:	
Storage:	-80 degrees
Date GC-MS Run:	2020-07-23
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:	RubenMF
Notes:	
Data:	https://doi.org/10.5281/zenodo.5810478