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G418 EvoPure®

Working Concentrations and G418 Selection Protocols

The following concentrations of active ingredient may be used as a guide for selection: mammalian cells
(400-1000μg/mL), plants (10-50 μg/mL), yeasts (500-1500 μg/mL), and dictyostelium (10-20μg/mL).
Cells have variable resistance and multiplying cells will be affected sooner than those that are not.

Cells in log phase may require three to seven days for selection. Varying concentrations should
be tested. Additionally, fibroblast contamination in mixed cultures may be suppressed by
using 100 μg/mL of the active ingredient for 2 days. Due to the higher purity of EvoPure®
agents, suggested G418 EvoPure™ selection concentrations are lower than normal G418.
For more rapid and accurate selections and to avoid tedious re-optimizations,
even higher concentrations of G418 can be used.

Species Tissue Cell Line Plasmid/ Vector Culture/Media Selection Concentration (μg/mL)
Normal G418 EvoPure® G418 EvoPure® G418 (rapid selection)
N/A N/A PhoeNX pcDNA3.1 DMEM, 10% FBS 500 450 700
Mouse Embryo mouse R1 ES pEGFP-N1 DMEM, 15% FBS, 2 mm L-glutamine, 50 μM β-mercaptoethanol, streptomycin-penicillin, sodium pyruvate, nonessential amino acids, and 10 ng/ml recombinant LIF 400 360 500
Mouse Subcutis L pCAGGS (constructed from pUC13) DMEM, 10% FBS 800 720 1000
Chinese hamster Ovary CHO pCAGGS (constructed from pUC13) DMEM with ribonucleosides and deoxyribonucleosides, 10% FBS 800 720 1000
Human Glioma U-138 MG pcDNA-CCRK, pcDNAT161A, pcDNA3.1 MEM, 10%FBS 500 450 700
Human Dermal NIH 3T3 p8-CAT DMEM, 10% FBS, 2mM glutamine 800 720 1000
Chinese hamster Ovary CHO-K1 pLc-eYFP and/or pHc-eGFP DMEM/F12, 10%FBS, 1.25%DMSO 400 360 600
Mouse Subcutis L pcdef3 Higuchi mediium, 10%FBS 700 630 900
Human Melanoma UACC903, pcDNA3 RPMI 1640, 10%FBS, 2 mM L-glutamine 600 540 800
UACC903(+6),
and SRS3
Sprague-Dawley rat vascular smooth muscle VSMCs pSPORT DMEM, 10% FBS 400 360 800
Human Skin Fibroblast pBABE and pMK DMEM, 4.5 g/l glucose, 110 μg/ml pyruvate, 5% FBS, 50 μg/ml uridine 200 180 400
Chicken B lymphoma DT40 modified pANMerCreMer, pAID-ctrl RPMI 1640, 10% FBS, 1% chicken serum, 50 μM 2-mercaptoethanol, 2 mM glutamine, 1 mM pyruvic acid 2000 1800 4000
Human Hepatocellular carcinoma SMMC-7721 pcDNA3.1(-) DMEM, 10% FBS 750 680 1200
Mouse Embryo NIH3T3 pBasic2 DMEM, 10% heat-inactivated FBS 1000 900 2000
Procyclic trypanosomes N/A Trypanosoma brucei 427-60 pI-ble-KO SDM-79 20 10 40
Mouse Embryo J1 ES pBS524 N/A 350 280 700
Human Kidney 293 pCR2.1-TOPO, pCMVSPORTbGal DMEM, 9% FBS 350 280 700
Chinese hamster Ovary CHO DUK- pcDNA3:VHCα2m(1), pcDNA3:VLCk α-minimal essential medium with 10% FBS, 10 mM Hepes 500 450 1000
Leishmania donovani N/A L. donovani promastigotes pX63-NEO, pX63-HYG DME-L 16 8 30
Mouse Bone marrow Ba/F3-6 pEFneo RPMI 1640, 10% FBS, 50 μM 2-mercaptoethanol 2000 1800 4000
Homo sapiens (human) Colon HCT-116 RFP RPMI 1640,10% FBS,2 mmol/L glutamine,37°C 50 30 100
Mus musculus (mouse)   PT67 pLEIN DMEM,10% heat-inactivated FBS 500-2000 450-1800 800-4000
Homo sapiens (human) Melanoma WM793 ptTA-neo   400 360 800
Mus musculus (mouse) Ascites RAW 264.7 pBK/CMV△p53asn RPMI 1640,10% (v/v) heat-inactivated
fetal-calf serum
400 360 800
Mus musculus (mouse) Neural crest SCTM41(Schwann cell) pcDNA 3.1 DMEM,10% FBS 800 720 1000
Homo sapiens (human) Lung BEAS-2B pHA-Jenv LHC-8 medium 1000 900 1500
Mus musculus (mouse)   EB3 Linearized knock-in vector Glasgow minimum essential medium,10% FCS,2-mercaptoethanol,LIF 150-200 100-180 300-500
Rat   MR-T1 pSV2-neo RPMI 1640,10% FBS,5%CO2,37°C 500 450 800
Homo sapiens (human) Bone hFOB pAR-HYG DMEM,10%(v/v) csFBS 300 270 500

Microbiological Spectrum:

Microorganism Effective Range
Concentrations (μg/ml)
Enterobacter 1-64
Enterococcus 8-32
Escherichia coli 0.5-64
Klebsiella 1-32
Pseudomonas aeruginosa 1-64
Salmonella 1-8
Serratia 1-8
Staphylococcus 0.5-4