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Jianshi Biotechnology (Beijing) Co., Ltd.

Plasmid DNA Mini-Prep Kit (TD429)

Item number	Product Name	Packaging Specifications	Price
TD429-100	Plasmid DNA Mini-Prep Kit	100 times/box	100
TD429-200	Plasmid DNA Mini-Prep Kit	200 times/box	180

Keywords

Jianshi Biomass Plasmid DNA Mini-Prep Kit

Product Overview

This kit is a classic small-scale plasmid DNA extraction kit (Catalog Nos. TD429-100/TD429-200), with the following key advantages:
※ Unique colorimetric reaction : P1 (red), P2 (blue), P3 (yellow); the color changes allow direct visualization of cell lysis and neutralization, making operational assessment intuitive and straightforward.
※ Fast, safe, and non-toxic : No toxic reagents such as phenol or chloroform are required, nor is ethanol precipitation necessary; the procedure is mild, ensuring greater safety for laboratory personnel.
※ High yield and good purity : The classic version can process up to 4 mL of bacterial lysate, with a single‑run yield of up to 100 µg, endotoxin levels ≤1 EU/µg, an A260/A280 ratio ≥1.8, and an A260/A230 ratio ≥2.0;
※ Directly used in downstream experiments : Purified plasmid DNA can be directly used in a variety of molecular biology experiments, including restriction digestion, transformation, PCR, in vitro transcription, sequencing, and transfection;
※ For research use only (RUO), not for clinical diagnostic use.

Product Features

※ Three-color colorimetric indicator system : P1 is red, P2 is blue, and P3 is yellow; after lysis, the solution becomes clear and viscous, and upon neutralization, a pale yellow flocculent precipitate forms, making the experimental status immediately apparent.
※ Remove organic solvents and ethanol precipitation. : The entire process employs column-based purification, eliminating the need for phenol/chloroform extraction and ethanol precipitation, thereby streamlining the workflow and minimizing exposure to hazardous substances.
※ Ultra-low endotoxin levels : In the classic version, endotoxin levels are ≤1 EU/µg, meeting the requirements for transfection-grade plasmids and making it particularly suitable for cell-based experiments that demand high purity.
※ High loading capacity, high recovery rate : The maximum binding capacity of a single column is 100 µg. For the standard version, it is recommended to process ≤4 ml of bacterial culture (OD ≈ 2–3), yielding up to 100 µg.
※ Compatible with a variety of downstream applications. : The purified product is free of protein, RNA, and genomic DNA contamination and can be directly used for restriction digestion, ligation, transformation, PCR, sequencing, transfection, and other applications.
※ Quick and easy to operate : Operates at room temperature, takes approximately 30 minutes from start to finish, requires no specialized equipment, and can be performed using either centrifugation or vacuum filtration.
※ Two specifications available : TD429-100 (100 cycles) and TD429-200 (200 cycles), catering to varying flux requirements;
※ Complete set of accompanying reagents The kit includes RNase A, P1/P2/P3 buffers, binding buffer, wash buffer 1/2, elution buffer, a purification column, and collection tubes; it is ready to use upon opening.
※ Strict quality control Each batch undergoes testing for potency, purity, and endotoxin levels, with a certificate of analysis provided to ensure consistent and reliable results.
※ One-year quality guarantee : A one-year warranty is provided from the date of purchase, and all reagents have undergone rigorous testing.

Product Composition and Technical Specifications (Classic Version)

Component / Parameter	TD429-100 (100 times)	TD429-200 (200 times)	Storage conditions
Classic version throughput	≤20 ml bacterial suspension	≤20 ml bacterial suspension	—
Classic version production volume	200 µg	200 µg	—
Endotoxin levels	≤0.1 EU/µg	≤0.1 EU/µg	—
RNase A solution	0.3 ml	0.6 ml	4°C
P1 (Red)	30 ml	60 ml	4°C
P2 (Blue)	30 ml	60 ml	Room temperature
P3 (Yellow)	30 ml	60 ml	Room temperature
Plasmid DNA binding solution	30 ml	60 ml	Room temperature
Plasmid DNA Wash Buffer 1	2 × 40 ml	4 × 40 ml	Room temperature
Plasmid DNA Wash Buffer 2	2 × 23 ml (Add 88 ml of 95% ethanol before use)	4 × 23 ml (Add 88 ml of 95% ethanol before use)	Room temperature
Plasmid DNA elution buffer	10 ml	20 ml	Room temperature
No. 3 PN purification column	100 pieces	200 pieces	Room temperature
2 ml collection tube	100 pieces	200 pieces	Room temperature
Operating temperature	Room temperature (15–30°C)		—
Operation Time	About 30 minutes		—
Plasmid DNA size	Up to 25 KB		—
Elution volume	≥50 µl (100 µl recommended)		—

Note: The “classic” protocol refers to an optimized procedure that processes ≤20 mL of bacterial culture, yields 200 µg of plasmid DNA, and achieves an endotoxin level of ≤0.1 EU/µg. Actual yield is influenced by factors such as plasmid copy number and the optical density (OD) of the bacterial culture.

Core Features

Three-color chromogenic system : P1 red → P2 blue → P3 yellow; the cleavage and neutralization states are immediately apparent.
Solvent-free : No phenol, chloroform, or ethanol precipitation required—safe and fast.
High purity : Abs260/280 ≥ 1.8, Abs260/230 ≥ 2.0
Low endotoxin : Endotoxin content < 1 EU/μg

Product Specifications

Item number : TD429-100 (100 cycles) / TD429-200 (200 cycles)
Processing capacity : 1–4 mL overnight bacterial culture (OD ≈ 2–3)
Column connection capacity : Maximum 100 μg
Elution volume : ≥50 μl (100 μl recommended)

Save and Operate

P1+RNase A : Store at 2–8°C
P2/P3/Binding Solution/Washing Solution : Store at room temperature
Operating temperature : Room temperature (15–30°C)
Operation Time : About 30 minutes

Plasmid DNA Mini Prep Kit – High-Frequency FAQ

1: What are the primary applications and key advantages of this kit?

It is used for the rapid extraction of plasmid DNA from Escherichia coli, requiring no toxic reagents such as phenol or chloroform throughout the entire process, nor does it necessitate ethanol precipitation. Its unique three‑color visualization system (P1 red, P2 blue, P3 yellow) enables intuitive monitoring of lysis and neutralization, ensuring simple and safe operation.

• The purified plasmid DNA exhibits exceptionally high purity (A260/280 ≥ 1.8, A260/230 ≥ 2.0) and an endotoxin level of less than 1 EU/μg, making it suitable for direct use in downstream applications such as restriction digestion, transformation, PCR, in vitro transcription, sequencing, and transfection.

• For research use only (RUO).

2: How does the three-color colorimetric system work? How can you determine whether lysis and neutralization have been completed?

P1 is a red suspension; upon addition of the bacterial suspension, it turns reddish‑turbid. After adding the blue reagent P2, the solution changes to a bluish‑purple color and, when lysis is complete, becomes a clear, viscous, bluish‑black liquid. Following neutralization with the yellow reagent P3, a pale yellow flocculent precipitate immediately forms, and the solution as a whole turns pale yellow, indicating that neutralization is complete.

• If, after neutralization, blue areas or clumping persist, it indicates insufficient neutralization; continue gently inverting the mixture until a uniform pale yellow color is achieved.

• Color changes are intuitive and reliable, eliminating the need for subjective judgment and significantly reducing operational complexity.

3: Why does this kit not require phenol, chloroform, or ethanol precipitation? How safe is it?

• The silica‑gel column purification method is employed: plasmid DNA specifically binds to the purification column, while impurities such as proteins, RNA, and genomic DNA are removed by washing buffers, followed by elution with a low‑salt buffer or water.

• The entire process avoids the use of any organic solvents, thereby eliminating the toxicity and flammability risks associated with phenol/chloroform, while also obviating the ethanol precipitation step, thus shortening the procedure and minimizing sample loss.

Safety Tips Although the reagent is non‑toxic, it is still recommended to wear gloves and safety goggles to prevent splashes from entering the eyes or mouth.

4: What is the quality of the purified plasmid DNA? Can it be used directly in demanding applications such as transfection?

The purified product exhibits exceptionally high purity: Abs260/280 ≥ 1.8, Abs260/230 ≥ 2.0, endotoxin content < 1 EU/μg, and no detectable residues of protein, RNA, or genomic DNA. It is suitable for direct use in enzymatic digestion, transformation, PCR, in vitro transcription, sequencing, and cell transfection assays.

• For transfection experiments, it is recommended to elute using elution buffer or sterile water, ensuring that the elution buffer is free from DNase/RNase contamination.

• If higher endotoxin specifications are required (e.g., for primary cell transfection), an endotoxin removal step can be incorporated, or an endotoxin‑free version can be selected.

5: What is the maximum volume of bacterial suspension that can be processed at one time? What is the maximum yield?

We recommend processing 1–4 mL of overnight‑grown bacterial culture (OD600 ≈ 2–3). The maximum binding capacity of a single purification column is 100 μg. The actual yield depends on factors such as plasmid copy number and the growth state of the strain; high‑copy plasmids typically yield 20–100 μg.

• If the OD value of the bacterial suspension is low (<2), you may appropriately increase the volume of the suspension, but the total cell load should not exceed the column’s maximum capacity.

• For higher yields, multiple purification columns can be used in parallel and their eluates pooled, or a high‑volume extraction kit can be selected.

6: What should I do if white precipitate or crystals appear in solution P2?

At lower ambient temperatures, SDS in P2 may precipitate. Place P2 in a 37°C water bath for a few minutes and gently invert it to dissolve; use only after the solution has become clear again. Avoid vigorous shaking to prevent excessive foaming, which could compromise lysis efficiency.

• Precipitation does not affect the reagent’s quality; after heating to dissolve, it can be used as usual.

• After use, immediately replace the cap to prevent evaporation and maintain the solution’s concentration.

7: What are the key precautions to observe during the procedure?

① Ensure the bacterial suspension is thoroughly resuspended, with no visible clumps; ② After adding P2, gently invert the tube 8 times—do not vortex—to prevent genomic DNA fragmentation; ③ Incubate for no more than 5 minutes; ④ Immediately invert the tube after adding P3 to neutralize, resulting in a uniform pale‑yellow flocculent precipitate; ⑤ When transferring the supernatant after centrifugation, avoid aspirating any precipitate; ⑥ Before use, add 88 mL of 95% ethanol to Washing Solution 2 as instructed and label the container accordingly.

• If the bacterial concentration is high (OD > 3), the volumes of P1, P2, P3, and the binding solution can be increased proportionally.

• Spin the column empty for 2 minutes before elution to remove residual ethanol and prevent interference with downstream reactions.

8: Supports both centrifugal and negative-pressure methods—how should you choose?

Both methods achieve comparable purification results. The centrifugation method is suitable for most laboratories and requires no additional equipment, whereas the vacuum‑assisted method is better suited for high‑throughput processing—when used with a vacuum manifold—and can handle multiple samples simultaneously at a faster pace.

• During negative-pressure operation, ensure that the purification column is tightly connected to the manifold to prevent air leaks; apply negative pressure at each step until the liquid has completely passed through.

• Centrifugation procedure: Centrifuge at 10,000×g for 1 minute each time; repeat the washing step twice.

9: How can the elution efficiency of plasmid DNA be improved?

Preheat the elution buffer (plasmid DNA elution buffer or sterile water) to 65–70°C. After adding it to the purification column matrix, let it stand at room temperature for 2 minutes, then centrifuge to elute. Preheating can improve elution efficiency by approximately 20–30%.

• For high‑concentration plasmid preparations, the elution volume can be reduced to 50 μl (we do not recommend going below 50 μl, as this may lower the recovery yield).

• The elution buffer should have a pH of 7.0–8.5; deionized water must be free of DNase.

10: What components are included in the kit? How should each component be stored?

TD429-100 (100 reactions) includes: RNase A solution, 0.3 ml (4°C); P1 (red), 30 ml (4°C); P2 (blue), 30 ml (room temperature); P3 (yellow), 30 ml (room temperature); plasmid DNA binding buffer, 30 ml (room temperature); plasmid DNA wash buffer 1, 2 × 40 ml (room temperature); plasmid DNA wash buffer 2, 2 × 23 ml (room temperature; add ethanol for first use); plasmid DNA elution buffer, 10 ml (room temperature); No. 3 PN purification columns, 100 units (room temperature); and 2‑ml collection tubes, 100 units (room temperature).

• Before first use, add all of the RNase A to P1, mix well, and store at 4°C.

• For each bottle of Wash Solution 2 (23 mL), add 88 mL of 95% ethanol, mix thoroughly, and mark the bottle to prevent duplicate additions.

• All reagents are covered by a one-year quality guarantee from the date of purchase.