説明
The HieffTM Endo-free Plasmid Maxi Kit V2 is designed for the rapid extraction of 0.5-4 mg of pure, high-copy plasmid DNA from 50-200 mL of culture broth (LB medium: 100-200 mL; 2YT medium: 50-100 mL), with an extraction efficiency of up to 80-90%. It offers high efficiency, speed, and convenience. The centrifugal adsorption column utilizes a unique silicon-based material developed by our company, combined with a proprietary lysis buffer formulation that maximizes the recovery of high-purity DNA. Paired with push filters, this kit simplifies the operation and ensures high DNA purity with low endotoxin levels (<1 EU/μg DNA). The extracted DNA is suitable for standard cell transfection and can be directly used in various molecular biology experiments such as restriction enzyme digestion, transformation, PCR, in vitro transcription, and sequencing.
Features
- High Binding Capacity – Silica membrane columns can bind up to 4 mg of plasmid DNA.
- High Purity & Low Endotoxin – Plasmid endotoxin levels <1 EU/μg (for mid-to-high copy plasmids), significantly lower than conventional methods (>2.5 EU/μg).
- Fast Workflow – Endotoxin removal and plasmid binding occur in a single step; entire process takes ~40 minutes.
- Flexible Compatibility – Compatible with both high-speed angled rotors and high-throughput horizontal rotors.
- Easy Operation – Push-filter design eliminates the need for careful supernatant handling.
Components
|
Category |
Components No. |
Name |
19037ES02 |
19037ES10 |
19037ES20 |
|
Part I |
19037-A |
RNase A (10 mg/mL) |
250 µL |
1200 µL |
1200 µL×2 |
|
Part II |
19037-B |
Binding Column E2 (HieffTM DNA Column E2) |
2 |
10 |
10×2 |
|
19037-C |
50 mL Collection Tube |
2 |
10 |
10×2 |
|
|
19037-D |
Push Filter |
2 |
10 |
10×2 |
|
|
19037-E |
Equilibration Buffer BS (Buffer BS)* |
10 mL |
50 mL |
100 mL |
|
|
19037-F |
Solution SI (Solution I) |
25 mL |
120 mL |
240 mL |
|
|
19037-G |
Solution SII (Solution II) |
25 mL |
120 mL |
240 mL |
|
|
19037-H |
Solution SIII (Solution III) |
20 mL |
100 mL |
200 mL |
|
|
19037-I |
Binding Solution PB (Solution PB)* |
7 mL |
32 mL |
64 mL |
|
|
19037-J |
Solution W1 (Solution W1) |
20 mL |
100 mL |
200 mL |
|
|
19037-K |
Wash Solution WS (concentrate)* |
12 mL |
48 mL |
48 mL x 2 |
|
|
19037-L |
Elution Buffer EB (Elution Buffer) |
10 mL |
30 mL |
60 mL |
|
|
19037-M |
Blue Buffer |
200 µL |
600 µL |
1200 µL |
Storage
Part I component 19037-A (RNase A, 10 mg/mL) should be stored at -25°C ~ -15°C, valid for 18 months.
Part II components should be stored at room temperature, valid for 18 months.
Application
Plasmid Extraction; Endo-free Plasmid Extraction.
Figures
1. Higher Yield
A 1200 mL bacterial culture was divided into six 200 mL aliquots and extracted using supplier A*(simplified endotoxin removal version), supplier B* and 19037ES kits. Each extraction was eluted twice with 1.2 mL per elution, and the eluates were collected in 1.5 mL centrifuge tubes. Elution volumes were accurately determined by weighing. Recovered plasmids were quantified using Nanodrop, analyzed by agarose gel electrophoresis, and tested for endotoxin levels.
2. Lower Endotoxin Levels
Figure 1. Plasmid extraction performance comparison.
(A) 19037 shows the lowest endotoxin levels; supplier B* lacks endotoxin removal, resulting in high residues.
(B) Five-fold diluted plasmids (3 μL each) were analyzed by agarose gel electrophoresis. The 19037 sample shows the brightest bands, consistent with Nanodrop concentrations, high supercoiled plasmid proportion, and no detectable RNA contamination.
Documents:
Safety Data Sheet
Manuals
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