Assay:	_A_MeUppS-EcoliExpr
Short Name:	MeUppS-EcoliExpr
Assay Class:	WET
Assay Type:	EcoliExpr
Title:	Candidate sequence from Moranella endobia
Description:	Expression in E. coli and activity testing
pISA Assay creation date:	2020-07-01
pISA Assay creator:	IG
Candidate sequence gbk file (file path or link):	_S_P4_expressionEcoli/reports/sequences/pMAL-c5XMeUppS.gb
Expression plasmid:	pMAL-c5X
Host E coli strain:	BL21(DE3)pLysS
Expression temperature:	30
Date of sample collection:	2020-07-02
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:	2020-07-02
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:	2020-07-03
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
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: