Efficient gene delivery is the cornerstone of modern molecular biology, from basic research to advanced therapies. But if you’ve ever struggled with low transfection efficiency—especially in sensitive cells like stem cells—the culprit may not be your transfection reagent, but your plasmid DNA.
Traditional plasmid prep methods often leave behind high levels of endotoxin (commonly >5 EU/µg). Even trace amounts can trigger apoptosis, stress responses, or misleading positive signals, compromising both research outcomes and therapeutic applications. For high-value workflows such as stem cell transfection, viral vector packaging, and gene therapy, endotoxin-free plasmid DNA isn’t just preferred—it’s essential.
Why Endotoxin-Free Matters ?
- Protects sensitive cells: Stem cells and primary cells are particularly vulnerable to endotoxin contamination, which can drastically lower transfection efficiency.
- Ensures reproducibility: Reducing endotoxin variation helps deliver consistent results across experiments.
- Meets clinical research needs: As gene therapy and vaccine development advance, the demand for large-scale, ultrapure plasmid DNA continues to rise.
Setting a New Standard in Plasmid Prep
With yields up to 4 mg and endotoxin levels consistently below 0.01 EU/µg, the Hieff™ Pro Endo-free Plasmid Maxi Kit sets a new benchmark for “high yield + ultrapure” plasmid preparation. Whether you’re tackling complex stem cell projects or building reliable vectors for gene therapy, this kit gives you the DNA quality you need to succeed.
Key Highlights:
High yield: Up to 4 mg plasmid DNA per prep from 50–200 mL culture; recovery efficiency 80–90%.
Ultra-pure: DNA with A260/280 ≥1.8, RNA-free, ready for downstream applications.
Lowest endotoxin levels: <0.01 EU/µg, ideal for sensitive cell types and therapeutic workflows.
Convenient workflow: Push-column filter design eliminates tedious supernatant handling and ensures cleaner DNA.
Broad Applications
Stem cell transfection – Improve efficiency in fragile, high-value cell systems.
Viral vector production – Ensure high-titer, reproducible lentivirus and AAV packaging.
Gene therapy research – Deliver GMP-like quality DNA for preclinical and translational studies.
Molecular biology workflows – Enzyme digestion, PCR, in vitro transcription, sequencing, and more.
Product Performance
- Higher Plasmid Yield and Purity
Equal volumes of bacterial culture (100 mL) were divided and extracted using Supplier A* and Yeasen Hieff™ Pro Endo-free Plasmid Maxi Kit (Cat#19038ES). Plasmid DNA was eluted twice with 1.2 mL each time. The exact elution volume was measured by weight. Plasmid yield and purity were analyzed by Nanodrop, band integrity confirmed by agarose gel electrophoresis, and endotoxin levels measured with the Shark Assay.
|
Kit |
Concentration (ng/µL) |
First Elution Volume (mL) |
Yield 1st Elution (µg) |
Yield 2nd Elution (µg) |
1st Elution Recovery Rate |
A260/280 |
A260/230 |
|
Supplier A* |
853.0 |
1.03 |
879 |
1068 |
82.30% |
1.81 |
2.29 |
|
Supplier A* |
631.6 |
1.00 |
630 |
739 |
85.24% |
1.81 |
2.26 |
|
19038ES |
942.9 |
1.07 |
1005 |
1189 |
84.53% |
1.84 |
2.41 |
|
19038ES |
928.8 |
1.06 |
983 |
1161 |
84.66% |
1.84 |
2.41 |
Result: The Hieff™ Pro Endo-free Plasmid Maxi Kit (19038ES) achieved 1.26× higher plasmid yield compared with Supplier A*, while maintaining comparable purity.
Lower Endotoxin Residual
|
Kit |
Endotoxin Residual |
|
Supplier A* |
~0.1 EU/µg |
|
Supplier A* |
~0.1 EU/µg |
|
19038ES |
<0.01 EU/µg |
|
19038ES |
<0.01 EU/µg |
Result: The 19038ES kit consistently delivered plasmid DNA with endotoxin levels below 0.01 EU/µg, significantly lower than Supplier A*.
Customer Feedback
From 200 mL bacterial culture, up to 1.8 mg plasmid DNA was recovered with excellent purity and ultra-low endotoxin, fully meeting daily transfection needs.
User-friendly workflow – the push-column filter allows easy removal of bacterial debris with a single pull.
Plasmid Yield and Endotoxin Comparison with Supplier B*
|
Kit |
Concentration (ng/µL) |
Yield (µg) |
A260/280 |
A260/230 |
Endotoxin Residual |
|
Supplier B* |
943.5 |
1415.3 |
1.83 |
2.21 |
~0.5 EU/µg |
|
Supplier B* |
852.5 |
1278.8 |
1.84 |
2.23 |
~0.5 EU/µg |
|
19038ES |
1221.4 |
1832.0 |
1.82 |
2.33 |
<0.1 EU/µg |
|
19038ES |
1264.1 |
1896.2 |
1.82 |
2.30 |
<0.1 EU/µg |
Result: Compared to Supplier B*, the 19038ES kit yielded higher DNA concentrations and lower endotoxin levels (<0.1 EU/µg vs ~0.5 EU/µg), ensuring superior plasmid quality for sensitive applications such as stem cell transfection and viral packaging.
FAQ – Endotoxin-Free Plasmid DNA Preparation
|
Question |
Answer |
|
What is the DNA yield of the Hieff™ Pro Endo-free Plasmid Maxi Kit? |
Up to 0.5–4 mg plasmid DNA can be obtained from 50–200 mL of high-copy bacterial culture, with recovery rates of 80–90%. |
|
How low are the endotoxin levels in the final DNA? |
Endotoxin levels are consistently controlled below 0.01 EU/µg DNA, making it suitable for sensitive cells like stem cells. |
|
What purity can I expect? |
DNA typically has an A260/280 ratio ≥1.8, is free of RNA, and ready for downstream applications such as sequencing, PCR, in vitro transcription, or enzymatic digestion. |
|
Is the kit compatible with both high-copy and low-copy plasmids? |
It is optimized for high-copy plasmids; low-copy plasmids may yield less DNA but can still be processed. |
|
What applications require endotoxin-free plasmids? |
Stem cell transfection, viral vector packaging (lentivirus, AAV), gene therapy studies, vaccine development, and any workflow sensitive to endotoxin contamination. |
|
How does this kit improve stem cell transfection? |
Endotoxin-free plasmids reduce cytotoxicity and false-positive signals, improving transfection efficiency and reproducibility in fragile cell types. |
|
How does the workflow differ from a standard maxi prep? |
The kit uses a triple endotoxin removal system and a push-column filter, eliminating tedious supernatant handling and ensuring ultra-clean DNA. |
|
Can the plasmid DNA be used directly for viral vector production? |
Yes. The endotoxin-free DNA quality is suitable for viral packaging workflows without additional purification. |
|
How does it compare to traditional plasmid extraction methods? |
Conventional methods often leave >5 EU/µg endotoxin and yield <1 mg DNA per prep, while this kit achieves up to 4 mg DNA with <0.01 EU/µg endotoxin. |
|
What culture volume should I use? |
The kit is designed for 50–200 mL bacterial cultures; using the recommended range ensures optimal yield and purity. |
Product list
|
Name |
Cat. No. |
Size |
|
19037ES02/10 |
2 T/10 T |
|
|
Hieff™ Pro Endo-free Plasmid Maxi Kit V2 |
19038ES02/10 |
2 T/10 T |