_A_Pc2704-EcoliExpr

Short Name: Pc2704-EcoliExpr Assay Class: WET Assay Type: EcoliExpr Title: Candidate sequence from P. citri Pc2704 Description: Expression in E. coli and activity testing of Pc2704 pISA Assay creation date: 2019-06-05 pISA Assay creator: IG Candidate sequence gbk file (file path or link): _S_P4_expressionEcoli/reports/sequences/pMAL-c5XPc2704.gb Expression plasmid: pMAL-c5X Host E coli strain: BL21(DE3)pLysS Expression temperature: 30C Date of sample collection: 2019-06-06 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: 2019-06-06 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; DMAPP 3 mM Date of LC-MS Run: 2019-06-07 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 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: