説明
HieffTM Endo-free Plasmid Mini Kit is designed for rapid extraction of 20–50 µg of high-purity plasmid DNA from 1–10 mL of E. coli LB culture. This kit features high efficiency, speed, and convenience. The spin column utilizes a proprietary silica-based matrix material developed by our company. Combined with our unique lysis buffer formulation, it enables maximum recovery of high-purity DNA. The extracted DNA has high purity and extremely low endotoxin residue (<0.1 EU/µg DNA), resulting in excellent performance in cell transfection. The DNA can be directly used in various molecular biology applications such as restriction digestion, transformation, PCR, in vitro transcription, and sequencing.
Feature
- Stable performance: Specially designed silica membranes ensure consistent DNA binding and reliable results.
- Fast and easy: Extract 20–50 µg high‑copy plasmid DNA from 1.5–5 mL E. coli LB culture in about 30 minutes.
- Pure DNA: Removes endotoxins to below 0.1 EU/µg DNA for excellent cell transfection.Specifications
Components
|
Category |
Components No. |
Name |
19021ES08 |
19021ES50 |
19021ES70 |
|
Part I |
19021-A |
RNase A (10 mg/mL) |
15 µL |
125 µL |
500 µL |
|
19021-B |
Endotoxin Removal Solution ER (ER Solution E2) |
500 µL |
5 mL |
20 mL |
|
|
Part II |
19021-C |
Buffer RS* (RS Buffer E2*) |
1.5 mL |
12.5 mL |
50 mL |
|
Part III |
19021-D |
Buffer AC (AC Buffer E2) |
500 µL |
5 mL |
20 mL |
|
19021-E |
DNA Binding Column E2 (HieffTM DNA Column E2) |
5 pcs |
50 pcs |
200 pcs |
|
|
19021-F |
2 mL Collection Tube (2 mL Collection Tube E2) |
5 pcs |
50 pcs |
200 pcs |
|
|
19021-G |
Lysis Buffer LB (LB Buffer E2) |
1.5 mL |
12.5 mL |
50 mL |
|
|
19021-H |
Binding Buffer BD (BD Buffer E2) |
1.5 mL |
12.5 mL |
50 mL |
|
|
19021-I |
Protein Lysis Buffer PL* (PL Buffer E2*) |
1.6 mL |
16 mL |
63 mL |
|
|
19021-J |
Wash Buffer W* (Wash Buffer*) |
1.3 mL |
13 mL |
50 mL |
|
|
19021-K |
Elution Buffer |
1 mL |
10 mL |
20 mL |
Shipping and Storage
Store Part I components at -25°C ~ -15°C; store Part II components at 2~8°C;
store Part III components at room temperature. The product is valid for 1 year.
Application
plasmid DNA Extraction; Endo-free Plasmid Extraction; Plasmid preparation for transfection.
Figures
1. Superior plasmid quality with ultra‑low endotoxin
Figure 1. Plasmid DNA extracted using Yeasen 19021, Supplier O*, and Supplier T* kits was transfected into HEK293 cells with pEGFP‑C1. Bright‑field and green‑fluorescence images were taken at 72 h post‑transfection, showing transfection efficiency in the order Yeasen 19021> Supplier T* > Supplier O*.
FAQ
Q: After adding the endotoxin removal reagent, did the solution not become cloudy?
A: It could be that the internal toxins were already low to begin with, or it could be that after adding the ER solution, due to differences in pH and inhibitors in the system, the aggregation state of the internal toxins reacted differently.
Q: Is the detection concentration of the non-toxic plasmid sample very high?
A: The customer used 20ml of the bacterial solution, which was too much. This resulted in insufficient activity of RNase.
Q: The concentration of the plasmid is low.
A: (1) For plasmids with a low copy number or larger than 10kb, the amount of bacteria used should be increased, and the subsequent buffer volume should also be proportionally increased; (2) The bacteria may not have been fully lysed.
Q: There is a precipitate in the hydrolysis solution.
A: The decomposition solution contains NaOH. There might be salt ions precipitated. It can be re-solubilized at 37℃ until the solution becomes clear before being used.
Q: A small band less than 100bp was observed in the agarose gel electrophoresis of the plasmid.
A: It could be that the RNA was not completely removed (perhaps due to too many samples or because the buffer containing RNase had reduced the activity of the RS enzyme), or that the lysis process was too prolonged, resulting in damage to the plasmids.
Q: The separation after removing endotoxins was not thorough enough.
A: Increase the centrifugal temperature; raise the centrifugal speed; extend the centrifugal time.
Q: The extraction yield is low.
A: Low-copy plasmids: For long fragments and expression-type vectors, the copy number is usually low. It is recommended to increase the amount of bacterial cells used and let the bacterial culture incubate overnight.
Bacterial strain issue: Plasmids were lost during the preservation of the bacterial strain. Before culturing, perform streaking to activate the bacteria, which will lead to a stable production output.
The bacterial cells were not fully lysed: Resuspend them thoroughly in the buffer to avoid clumping;
Reagent preparation error: The solution with precipitate formation needs to be heated; The volume of added ethanol is incorrect.
Insufficient elution: Preheat the elution solution and perform a second elution.
Q: There is genomic contamination.
A: The cultivation period is too long: The recommended cultivation time should be within 12 to 16 hours; the lysis is insufficient.
Q: The results at the downstream stage were not satisfactory.
A: Salt ion contamination; Ethanol contamination; Plasmid degradation; Elution of chromatography column membrane.
Documents:
Safety Data Sheet
Manuals
19021_Manual_Ver.EN20250806.pdf
Related Blog
Citations & References:
[1] Han P, Cao P, Yue J, et al. Knockdown of hnRNPA1 Promotes NSCLC Metastasis and EMT by Regulating Alternative Splicing of LAS1L exon 9. Front Oncol. 2022;12:837248. Published 2022 Jun 23. doi:10.3389/fonc.2022.837248(IF:5.738)
[2] Zhang J, Wang W, Feng N, Jiang X, Zhu S, Chen YQ. Ndufa6 regulates adipogenic differentiation via Scd1. Adipocyte. 2021;10(1):646-657. doi:10.1080/21623945.2021.2007590(IF:4.534)
[3] Zhu S, Zhang J, Wang W, Jiang X, Chen YQ. Blockage of NDUFB9-SCD1 pathway inhibits adipogenesis : Blockage of NDUFB9-SCD1 pathway inhibits adipogenesis. J Physiol Biochem. 2022;78(2):377-388. doi:10.1007/s13105-022-00876-7(IF:4.158)
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