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Experiment Overview

Repository ID: FR-FCM-ZZQM Experiment name: Heterogeneity and plasmid copy number of E. coli carrying pSEVA plasmids MIFlowCyt score: 87.94%
Primary researcher: Michael Jahn PI/manager: Michael Jahn Uploaded by: Michael Jahn
Experiment dates: 2015-06-01 - 2015-12-31 Dataset uploaded: Apr 2016 Last updated: Nov 2016
Keywords: [reporter] [Fluorescent proteins] [EGFP] [plasmid] [plasmid copy number] [Heterologous expression] [Escherichia coli] [replication system] Manuscripts:
Organizations: Helmholtz-Centre for Environmental Research - UFZ, Env. microbiology, Leipzig, (Germany)
Purpose: The aim was to test performance and identify population heterogeneity in a set of standardized plasmids from the SEVA repository. The seven plasmids differed in their replication system, while the plasmid backbone containing lacIq/Ptrc and an EGFP reporter remained identical for all.
Conclusion: Six out of the seven constructs showed homogeneous distribution. Replication system 5 (pSEVA254-AEB, RSF1010 replication origin) showed a considerable amount of heterogeneity in terms of EGFP expression. Two exemplary populations were sorted (pSEVA244-AEB, pSEVA254-AEB) and the plasmid copy number determined on the sub-population level showed extreme variation. For details regarding study concept and sample description, see: Jahn et al., Microbial Cell Factories, 2016.
Comments: None
Funding: This work was supported by the European Union Regional Development Fund (ERDF) and the Free State of Saxony (Germany).
Quality control: Non-fluorescent control strains carrying EGFP-free plasmids were used as a control.


Experiment variables

Timepoints
· 0 ATH_02_Nov_15_08.fcs · ATH_02_Nov_15_09.fcs · ATH_02_Nov_15_10.fcs · ATH_02_Nov_15_11.fcs · ATH_02_Nov_15_12.fcs · ATH_02_Nov_15_13.fcs · ATH_02_Nov_15_14.fcs · AMJ_23_Jun_15_13.fcs · AMJ_23_Jun_15_14.fcs · AMJ_23_Jun_15_16.fcs · AMJ_23_Jun_15_17.fcs · AMJ_23_Jun_15_18.fcs · AMJ_23_Jun_15_19.fcs · ATH_03_Dec_15_08.fcs
· 4 ATH_02_Nov_15_15.fcs · ATH_02_Nov_15_16.fcs · ATH_02_Nov_15_17.fcs · ATH_02_Nov_15_18.fcs · ATH_02_Nov_15_19.fcs · ATH_02_Nov_15_20.fcs · ATH_02_Nov_15_21.fcs · AMJ_23_Jun_15_20.fcs · AMJ_23_Jun_15_21.fcs · AMJ_23_Jun_15_23.fcs · AMJ_23_Jun_15_24.fcs · AMJ_23_Jun_15_25.fcs · AMJ_23_Jun_15_26.fcs · ATH_03_Dec_15_12.fcs
· 8 ATH_02_Nov_15_22.fcs · ATH_02_Nov_15_23.fcs · ATH_02_Nov_15_24.fcs · ATH_02_Nov_15_25.fcs · ATH_02_Nov_15_26.fcs · ATH_02_Nov_15_27.fcs · ATH_02_Nov_15_28.fcs · AMJ_23_Jun_15_27.fcs · AMJ_23_Jun_15_28.fcs · AMJ_23_Jun_15_30.fcs · AMJ_23_Jun_15_31.fcs · AMJ_23_Jun_15_32.fcs · AMJ_23_Jun_15_33.fcs · ATH_03_Dec_15_16.fcs
· 24 ATH_02_Nov_15_29.fcs · ATH_02_Nov_15_30.fcs · ATH_02_Nov_15_31.fcs · ATH_02_Nov_15_32.fcs · ATH_02_Nov_15_33.fcs · ATH_02_Nov_15_34.fcs · ATH_02_Nov_15_35.fcs · AMJ_23_Jun_15_34.fcs · AMJ_23_Jun_15_35.fcs · AMJ_23_Jun_15_37.fcs · AMJ_23_Jun_15_38.fcs · AMJ_23_Jun_15_39.fcs · AMJ_23_Jun_15_40.fcs · ATH_03_Dec_15_20.fcs

Conditions
· p224-AEB ATH_02_Nov_15_08.fcs · ATH_02_Nov_15_15.fcs · ATH_02_Nov_15_22.fcs · ATH_02_Nov_15_29.fcs · AMJ_23_Jun_15_13.fcs · AMJ_23_Jun_15_20.fcs · AMJ_23_Jun_15_27.fcs · AMJ_23_Jun_15_34.fcs
· p234-AEB ATH_02_Nov_15_09.fcs · ATH_02_Nov_15_16.fcs · ATH_02_Nov_15_23.fcs · ATH_02_Nov_15_30.fcs · AMJ_23_Jun_15_14.fcs · AMJ_23_Jun_15_21.fcs · AMJ_23_Jun_15_28.fcs · AMJ_23_Jun_15_35.fcs
· p244-AEB ATH_02_Nov_15_10.fcs · ATH_02_Nov_15_17.fcs · ATH_02_Nov_15_24.fcs · ATH_02_Nov_15_31.fcs · ATH_03_Dec_15_08.fcs · ATH_03_Dec_15_12.fcs · ATH_03_Dec_15_16.fcs · ATH_03_Dec_15_20.fcs
· p254-AEB ATH_02_Nov_15_11.fcs · ATH_02_Nov_15_18.fcs · ATH_02_Nov_15_25.fcs · ATH_02_Nov_15_32.fcs · AMJ_23_Jun_15_16.fcs · AMJ_23_Jun_15_23.fcs · AMJ_23_Jun_15_30.fcs · AMJ_23_Jun_15_37.fcs
· p264-AEB ATH_02_Nov_15_12.fcs · ATH_02_Nov_15_19.fcs · ATH_02_Nov_15_26.fcs · ATH_02_Nov_15_33.fcs · AMJ_23_Jun_15_17.fcs · AMJ_23_Jun_15_24.fcs · AMJ_23_Jun_15_31.fcs · AMJ_23_Jun_15_38.fcs
· p274-AEB ATH_02_Nov_15_13.fcs · ATH_02_Nov_15_20.fcs · ATH_02_Nov_15_27.fcs · ATH_02_Nov_15_34.fcs · AMJ_23_Jun_15_18.fcs · AMJ_23_Jun_15_25.fcs · AMJ_23_Jun_15_32.fcs · AMJ_23_Jun_15_39.fcs
· p284-AEB ATH_02_Nov_15_14.fcs · ATH_02_Nov_15_21.fcs · ATH_02_Nov_15_28.fcs · ATH_02_Nov_15_35.fcs · AMJ_23_Jun_15_19.fcs · AMJ_23_Jun_15_26.fcs · AMJ_23_Jun_15_33.fcs · AMJ_23_Jun_15_40.fcs

Sample Type
· R1 ATH_02_Nov_15_08.fcs · ATH_02_Nov_15_09.fcs · ATH_02_Nov_15_10.fcs · ATH_02_Nov_15_11.fcs · ATH_02_Nov_15_12.fcs · ATH_02_Nov_15_13.fcs · ATH_02_Nov_15_14.fcs · ATH_02_Nov_15_15.fcs · ATH_02_Nov_15_16.fcs · ATH_02_Nov_15_17.fcs · ATH_02_Nov_15_18.fcs · ATH_02_Nov_15_19.fcs · ATH_02_Nov_15_20.fcs · ATH_02_Nov_15_21.fcs · ATH_02_Nov_15_22.fcs · ATH_02_Nov_15_23.fcs · ATH_02_Nov_15_24.fcs · ATH_02_Nov_15_25.fcs · ATH_02_Nov_15_26.fcs · ATH_02_Nov_15_27.fcs · ATH_02_Nov_15_28.fcs · ATH_02_Nov_15_29.fcs · ATH_02_Nov_15_30.fcs · ATH_02_Nov_15_31.fcs · ATH_02_Nov_15_32.fcs · ATH_02_Nov_15_33.fcs · ATH_02_Nov_15_34.fcs · ATH_02_Nov_15_35.fcs
· R2 AMJ_23_Jun_15_13.fcs · AMJ_23_Jun_15_14.fcs · AMJ_23_Jun_15_16.fcs · AMJ_23_Jun_15_17.fcs · AMJ_23_Jun_15_18.fcs · AMJ_23_Jun_15_19.fcs · AMJ_23_Jun_15_20.fcs · AMJ_23_Jun_15_21.fcs · AMJ_23_Jun_15_23.fcs · AMJ_23_Jun_15_24.fcs · AMJ_23_Jun_15_25.fcs · AMJ_23_Jun_15_26.fcs · AMJ_23_Jun_15_27.fcs · AMJ_23_Jun_15_28.fcs · AMJ_23_Jun_15_30.fcs · AMJ_23_Jun_15_31.fcs · AMJ_23_Jun_15_32.fcs · AMJ_23_Jun_15_33.fcs · AMJ_23_Jun_15_34.fcs · AMJ_23_Jun_15_35.fcs · AMJ_23_Jun_15_37.fcs · AMJ_23_Jun_15_38.fcs · AMJ_23_Jun_15_39.fcs · AMJ_23_Jun_15_40.fcs · ATH_03_Dec_15_08.fcs · ATH_03_Dec_15_12.fcs · ATH_03_Dec_15_16.fcs · ATH_03_Dec_15_20.fcs
· controls ATH_02_Nov_15_03.fcs · ATH_02_Nov_15_04.fcs · ATH_02_Nov_15_06.fcs · AMJ_23_Jun_15_03.fcs · AMJ_23_Jun_15_04.fcs · AMJ_23_Jun_15_06.fcs

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