_ASSAY_METADATA.TXT
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_p_SUSPHIRE/_I_T31_mealybug/_S_P3_expressionEcoli/_A_Trans57-EcoliExpr/

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Assay:	_A_Trans57-EcoliExpr
Short Name:	Trans57-EcoliExpr
Assay Class:	WET
Assay Type:	EcoliExpr
Title:	Candidate sequence from P. citri Trans57
Description:	Expression in E. coli and activity testing of PcTrans57
pISA Assay creation date:	2020-11-25
pISA Assay creator:	IG
Candidate sequence gbk file (file path or link):	_S_P4_expressionEcoli/reports/sequences/pMAL-c5Xtrans57.gb
Expression plasmid:	pMAL-c5X
Host E coli strain:	BL21(DE3)pLysS
Expression temperature:	30C
Date of sample collection:	2020-11-26
Sample purification protocol (file path, link or citation):	_S_P4_expressionEcoli/reports/methods/Purification of MALfused proteins.docx
Sample purification method:	Amilose resin
Date of activity assay:	2020-11-26
Activity assay buffer:	35 mM HEPES, pH 7.6; 10 mM MgCl2, 5 mM beta-mercaptoethanol
Activity assay incubation time:	overnight
Substrates names and concentrations:	IPP/DMAPP 100 mcM each; DMAPP 200 mcM
Date of LC-MS Run:	2020-11-27
LC instrument:	1260 Infinity (Agilent)
LC column:	Poroshell 120 EC-C18, 3.0 x 50 mm, 2.7 (Agilent)
LC mobile phase A:	5 mM ammonium bicarbonate
LC mobile phase B:	acetonitrile
Chromatography protocol (file path, link or citation):	_S_P4_expressionEcoli/reports/methods/LCMS_gradient1.docx
MS detector:	G6120B quadrupole mass spectrometry detector (Agilent)
MS ionization:	atmospheric pressure ionization - electrospray (API-ES)
MS mode:	negative SIM
MS mode SIM mz:	313; 381
LC-MS Software and version:	Agilent ChemStation Rev.C.01.07 SR2[255]
Mass spectrometry protocol (file path, link or citation):	_S_P4_expressionEcoli/reports/methods/MS.docx
Quantification protocol (file path, link or citation):	_S_P4_expressionEcoli/reports/methods/LCMS_calibration_GPP.xlsx
Lab manager:	IG
Operator:	IG
Notes:	
Phenodata:	None
Featuredata:	
Data:	
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Created: 16th May 2022 at 10:30

Last updated: 6th Nov 2024 at 15:25

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Version 1 (earliest) Created 16th May 2022 at 10:30 by Marko Petek

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