The E.Z.N.A.® Endo-Free Plasmid DNA Maxi Kit provides an efficient method for the large-scale isolation of low-endotoxin plasmid DNA. The system utilizes Lysate Clearance Filter Syringes instead of centrifugation to rapidly clear bacterial lysates post-alkaline lysis. Following lysate clearance, endotoxins are removed, and the protocol follows a simple “bind-wash-elute” procedure to deliver high-quality plasmid DNA. The yields vary according to plasmid copy number, E. coli strain, and growth conditions. 50-200 mL of bacterial cultures in LB medium typically produces 0.6-1.2 mg of high copy number plasmid DNA. Up to 500 mL of culture may be processed when working with low copy number plasmid. The system uses centrifugation or vacuum technology for plasmid purification and eliminates the time-consuming gravity-flow columns that require alcohol precipitation. Purified plasmid DNA is suitable for sensitive downstream applications such as transfection of mammalian cells, cloning, automated fluorescent DNA sequencing (typical reads exceed 800 bp), next-generation sequencing, restriction enzyme digestion, ligation, PCR, in vitro transcription, transformation, etc.
For Research Use Only. Not for use in diagnostic procedures.
| FEATURES | SPECIFICATIONS |
|---|---|
| Downstream Application | Transfection, gene therapy, cloning, in vitro Transcription, Next-Generation Sequencing, PCR, Transformation, restriction digestion, ligation |
| Starting material | 50-200 mL LB culture with OD600 between 2 and 3; or equivalent |
| Plasmid type | High-copy, low-copy, cosmid DNA |
| Processing mode | Manual (centrifugation or vacuum) |
| Throughput | 1 – 24 |
| DNA binding technology | Silica Maxi Spin Column |
| Lysate clearance method | Filtration using Lysate clearance syringe |
| Processing time | <45 minutes |
| Yield | 600-1200 µg for high copy-number; 50-300 µg for low copy-number |
| Special note | Includes endotoxin removal step |
High-Quality Plasmid DNA Purification
Figure 1. pGEM and pGL4.51 plasmids were purified from 200 mL DH5alpha cultures harboring the plasmids and eluted in 1500 µL volume using Omega Bio-tek’s E.Z.N.A.® Endo-Free Plasmid DNA Maxi Kit. Plasmid DNA concentrations were determined by optical density measurements using Thermo Scientific’s NanoDrop™ 2000c system. Charles River’s Endosafe LAL Cartridge 1.0 – 0.01 EU/mL was used with the Nexgen-PTS device. All samples have ideal A260/A280 (~1.8) and A260/A230 (~2.0) absorbance ratios, indicating high-quality purified DNA.
Unfragmented Plasmid Purification
Figure 2. 200ng purified pGEM and pGL4.51 plasmid were analyzed on a 0.8% Agarose gel. Plasmids were isolated from 50 mL DH5α cultures using Omega Bio-tek’s E.Z.N.A.® Endo-Free Plasmid DNA Maxi Kit. Plasmids were digested with HF-ScaI to linearize the plasmids.
Sequencing-Quality Plasmid Purification
| Plasmid Type | Sample | Trace Score | Continuous Read Length | QV20+ |
|---|---|---|---|---|
| pGEM | 1 | 57 | 952 | 959 |
| 2 | 57 | 955 | 962 | |
| 3 | 57 | 965 | 972 | |
| pGL4.51 | 1 | 58 | 962 | 970 |
| 2 | 56 | 962 | 968 | |
| 3 | 57 | 964 | 972 |
Table 1. pGEM and pGL4.51 plasmids were purified from 200 mL DH5alpha cultures and eluted in 1500 µL volume using Omega Bio-tek’s E.Z.N.A.® Endo-Free Plasmid DNA Maxi Kit. The plasmids were then used for Sanger sequencing. All samples display high continuous read length and QV20+ scores, indicating plasmids of high-quality isolated from this kit.
Product Information
Safety Data Sheets
| Components | Hazard Standards | Languages | Link | hf:tax:dlp_document_language | hf:tax:dlp_document_hazard-standard |
|---|---|---|---|---|---|
| DNA Wash Buffer | GHS | English | english | ghs | |
| DNA Wash Buffer | GHS | Spanish | spanish | ghs | |
| DNA Wash Buffer | REACH | English | english | reach | |
| DNA Wash Buffer | REACH | Danish | danish | reach | |
| DNA Wash Buffer | REACH | Finnish | finnish | reach | |
| DNA Wash Buffer | REACH | French | french | reach | |
| DNA Wash Buffer | REACH | German | german | reach | |
| DNA Wash Buffer | REACH | Italian | italian | reach | |
| DNA Wash Buffer | REACH | Norwegian | norwegian | reach | |
| DNA Wash Buffer | REACH | Spanish | spanish | reach | |
| DNA Wash Buffer | REACH | Swedish | swedish | reach | |
| DNA Wash Buffer | WHMS | English | english | whms | |
| DNA Wash Buffer | WHMS | French | french | whms | |
| DNA Wash Buffer | REACH | Latvian | latvian | reach | |
| DNA Wash Buffer | REACH | Polish | polish | reach | |
| EHBC Buffer | GHS | English | english | ghs | |
| EHBC Buffer | GHS | Spanish | spanish | ghs | |
| EHBC Buffer | REACH | English | english | reach | |
| EHBC Buffer | REACH | Danish | danish | reach | |
| EHBC Buffer | REACH | Finnish | finnish | reach | |
| EHBC Buffer | REACH | French | french | reach | |
| EHBC Buffer | REACH | German | german | reach | |
| EHBC Buffer | REACH | Italian | italian | reach | |
| EHBC Buffer | REACH | Norwegian | norwegian | reach | |
| EHBC Buffer | REACH | Spanish | spanish | reach | |
| EHBC Buffer | REACH | Swedish | swedish | reach | |
| EHBC Buffer | WHMS | English | english | whms | |
| EHBC Buffer | WHMS | French | french | whms | |
| EHBC Buffer | REACH | Latvian | latvian | reach | |
| EHBC Buffer | REACH | Polish | polish | reach | |
| Endo-Free Elution Buffer | GHS | English | english | ghs | |
| Endo-Free Elution Buffer | GHS | Spanish | spanish | ghs | |
| Endo-Free Elution Buffer | REACH | English | english | reach | |
| Endo-Free Elution Buffer | REACH | Danish | danish | reach | |
| Endo-Free Elution Buffer | REACH | Finnish | finnish | reach | |
| Endo-Free Elution Buffer | REACH | French | french | reach | |
| Endo-Free Elution Buffer | REACH | German | german | reach | |
| Endo-Free Elution Buffer | REACH | Italian | italian | reach | |
| Endo-Free Elution Buffer | REACH | Norwegian | norwegian | reach | |
| Endo-Free Elution Buffer | REACH | Spanish | spanish | reach | |
| Endo-Free Elution Buffer | REACH | Swedish | swedish | reach | |
| Endo-Free Elution Buffer | WHMS | English | english | whms | |
| Endo-Free Elution Buffer | WHMS | French | french | whms | |
| Endo-Free Elution Buffer | REACH | Latvian | latvian | reach | |
| Endo-Free Elution Buffer | REACH | Polish | polish | reach | |
| ETR Binding Buffer | GHS | English | english | ghs | |
| ETR Binding Buffer | GHS | Spanish | spanish | ghs | |
| ETR Binding Buffer | REACH | English | english | reach | |
| ETR Binding Buffer | REACH | Danish | danish | reach | |
| ETR Binding Buffer | REACH | Finnish | finnish | reach | |
| ETR Binding Buffer | REACH | French | french | reach | |
| ETR Binding Buffer | REACH | German | german | reach | |
| ETR Binding Buffer | REACH | Italian | italian | reach | |
| ETR Binding Buffer | REACH | Norwegian | norwegian | reach | |
| ETR Binding Buffer | REACH | Spanish | spanish | reach | |
| ETR Binding Buffer | REACH | Swedish | swedish | reach | |
| ETR Binding Buffer | WHMS | English | english | whms | |
| ETR Binding Buffer | WHMS | French | french | whms | |
| ETR Binding Buffer | REACH | Latvian | latvian | reach | |
| ETR Binding Buffer | REACH | Polish | polish | reach | |
| ETR Wash Buffer | GHS | English | english | ghs | |
| ETR Wash Buffer | GHS | Spanish | spanish | ghs | |
| ETR Wash Buffer | REACH | English | english | reach | |
| ETR Wash Buffer | REACH | Danish | danish | reach | |
| ETR Wash Buffer | REACH | Finnish | finnish | reach | |
| ETR Wash Buffer | REACH | French | french | reach | |
| ETR Wash Buffer | REACH | German | german | reach | |
| ETR Wash Buffer | REACH | Italian | italian | reach | |
| ETR Wash Buffer | REACH | Norwegian | norwegian | reach | |
| ETR Wash Buffer | REACH | Spanish | spanish | reach | |
| ETR Wash Buffer | REACH | Swedish | swedish | reach | |
| ETR Wash Buffer | WHMS | English | english | whms | |
| ETR Wash Buffer | WHMS | French | french | whms | |
| ETR Wash Buffer | REACH | Latvian | latvian | reach | |
| ETR Wash Buffer | REACH | Polish | polish | reach | |
| N3 Buffer | GHS | English | english | ghs | |
| N3 Buffer | GHS | Spanish | spanish | ghs | |
| N3 Buffer | REACH | English | english | reach | |
| N3 Buffer | REACH | Danish | danish | reach | |
| N3 Buffer | REACH | Finnish | finnish | reach | |
| N3 Buffer | REACH | French | french | reach | |
| N3 Buffer | REACH | German | german | reach | |
| N3 Buffer | REACH | Italian | italian | reach | |
| N3 Buffer | REACH | Norwegian | norwegian | reach | |
| N3 Buffer | REACH | Spanish | spanish | reach | |
| N3 Buffer | REACH | Swedish | swedish | reach | |
| N3 Buffer | WHMS | English | english | whms | |
| N3 Buffer | WHMS | French | french | whms | |
| N3 Buffer | REACH | Latvian | latvian | reach | |
| N3 Buffer | REACH | Polish | polish | reach | |
| RNase A | GHS | English | english | ghs | |
| RNase A | GHS | Spanish | spanish | ghs | |
| RNase A | REACH | English | english | reach | |
| RNase A | REACH | Danish | danish | reach | |
| RNase A | REACH | Finnish | finnish | reach | |
| RNase A | REACH | French | french | reach | |
| RNase A | REACH | German | german | reach | |
| RNase A | REACH | Italian | italian | reach | |
| RNase A | REACH | Norwegian | norwegian | reach | |
| RNase A | REACH | Spanish | spanish | reach | |
| RNase A | REACH | Swedish | swedish | reach | |
| RNase A | WHMS | English | english | whms | |
| RNase A | WHMS | French | french | whms | |
| RNase A | REACH | Latvian | latvian | reach | |
| RNase A | REACH | Polish | polish | reach | |
| Solution I | GHS | English | english | ghs | |
| Solution I | GHS | Spanish | spanish | ghs | |
| Solution I | REACH | English | english | reach | |
| Solution I | REACH | Danish | danish | reach | |
| Solution I | REACH | Finnish | finnish | reach | |
| Solution I | REACH | French | french | reach | |
| Solution I | REACH | German | german | reach | |
| Solution I | REACH | Italian | italian | reach | |
| Solution I | REACH | Norwegian | norwegian | reach | |
| Solution I | REACH | Spanish | spanish | reach | |
| Solution I | REACH | Swedish | swedish | reach | |
| Solution I | WHMS | English | english | whms | |
| Solution I | WHMS | French | french | whms | |
| Solution I | REACH | Latvian | latvian | reach | |
| Solution I | REACH | Polish | polish | reach | |
| Solution II | GHS | English | english | ghs | |
| Solution II | GHS | Spanish | spanish | ghs | |
| Solution II | REACH | English | english | reach | |
| Solution II | REACH | Danish | danish | reach | |
| Solution II | REACH | Finnish | finnish | reach | |
| Solution II | REACH | French | french | reach | |
| Solution II | REACH | German | german | reach | |
| Solution II | REACH | Italian | italian | reach | |
| Solution II | REACH | Norwegian | norwegian | reach | |
| Solution II | REACH | Spanish | spanish | reach | |
| Solution II | REACH | Swedish | swedish | reach | |
| Solution II | WHMS | English | english | whms | |
| Solution II | WHMS | French | french | whms | |
| Solution II | REACH | Latvian | latvian | reach | |
| Solution II | REACH | Polish | polish | reach |

