All

All
Product Management
News
Introduction

Language
- CN
- RU
- EN

Jianshi Biotechnology (Beijing) Co., Ltd.

Plasmid DNA Rapid Mini‑to‑Mid‑Scale Extraction Kit (TD435-CQ)

Item number	Product Name	Packaging Specifications	Price
TD435-CQ-50	Rapid Mini-Prep Kit for Plasmid DNA (Isopropanol Method)	50 times/box	260
TD435-CQ-100	Rapid Mini-Prep Kit for Plasmid DNA (Isopropanol Method)	100 times/box	500

Keywords

Jianshi Biomass Plasmid DNA Rapid Mini‑to‑Mid‑Volume Extraction Kit

Product Overview

This kit is a rapid mid‑scale plasmid DNA extraction kit (Catalog No. TD435, Q‑Series), with the following key advantages:
※ Unique colorimetric reaction : P1 (red), P2, and P4—color changes allow direct visualization of cell lysis and neutralization, making operation and interpretation straightforward and intuitive.
※ 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 : Each extraction yields approximately 100 µg of plasmid DNA, with extremely low endotoxin levels (≤0.5 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 sensitive molecular biology experiments, including restriction digestion, transformation, PCR, in vitro transcription, sequencing, transfection, and in vivo cell studies;
※ Quick and easy to operate : The entire process takes only 16 minutes, can be performed at room temperature, and supports both centrifugation and negative-pressure purification methods.
※ For research use only (RUO), not for clinical diagnostic use.

Product Features

※ Three-color colorimetric indicator system : P1 red, P2, and P4 turn a clear, bright pale pink upon hydrolysis; after neutralization, they shift to yellow and precipitate abundant white flocculent material, making the operational 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 chemicals.
※ Ultra-low endotoxin levels : Endotoxin content ≤ 0.5 EU/µg, meeting the requirements for transfection-grade plasmids and particularly suitable for cell-based experiments that are sensitive to purity;
※ High loading capacity, high recovery rate : Can process 5–15 mL of bacterial lysate per run (OD ≈ 3), with a yield of up to 100 µg; the column exhibits strong binding capacity.
※ Compatible with a variety of downstream applications. : The purified product is free of protein, RNA, and genomic DNA residues 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 16 minutes from start to finish, requires no specialized equipment, and can be performed using either centrifugation or vacuum filtration.
※ Two specifications available : TD435-CQ-50 (50 cycles), TD435-CQ-100 (100 cycles), meeting diverse throughput requirements;
※ Matching funnel and collection tube The kit includes a 25 mL funnel and a 2 mL collection tube, facilitating negative-pressure operation and high-volume sample loading.
※ Strict quality control Each batch undergoes testing for potency, purity, and endotoxin levels, with an accompanying quality‑control report 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 (All Models)

Component / Parameter	5 times	50 times	100 times	200 times	Storage conditions
RNase A solution	100 μl	600 μl	2×600 μl	4×600 μl	4°C
P1 (Red)	10 ml	60 ml	2×60 ml	4×60 ml	4°C
P2	10 ml	60 ml	2×60 ml	4×60 ml	Room temperature
P4	10 ml	60 ml	2×60 ml	4×60 ml	Room temperature
Endo-Free Plasmid DNA Wash Buffer	5 ml	30 ml	2×30 ml	3 × 30 ml	Room temperature (add ethanol upon first use)
Plasmid DNA elution solution	1 ml	5 ml	10 ml	2×10 ml	Room temperature
No. 3 PN Purification Column	5 pieces	50 pieces	2 × 50 pieces	4 × 50 pieces	Room temperature
25 ml funnel	10 pieces	50 pieces	100 pieces	200 pieces	Room temperature
2 mL collection tube	5 pieces	50 pieces	2 × 50 pieces	200 pieces	Room temperature
Product Features	• Plasmid DNA yield: approximately 100 μg per preparation (high-copy plasmid) • Purity: Abs260/280 ≥ 1.8, Abs260/230 ≥ 2.0 • Endotoxin: ≤0.5 EU/μg • Plasmid DNA size: up to 20 kb • Elution volume: ≥50 μl (100 μl recommended) • Operating temperature: Room temperature (15–30°C) • Operating time: approximately 16 minutes				—

Note: Actual yield is influenced by factors such as plasmid copy number and bacterial culture OD; endotoxin levels are correlated with the amount of bacteria processed. We recommend following the recommended loading volume specified in the protocol.

Core Features

Three-color chromogenic system : P1 red → P2 → P4; the cleavage and neutralization states are immediately apparent.
16-minute rapid extraction : Operates at room temperature; no phenol/chloroform/ethanol precipitation required.
High purity, low endotoxin : Abs260/280 ≥ 1.8, endotoxin ≤ 0.5 EU/μg
Yield approximately 100 μg. : Suitable for 5–15 mL of bacterial culture, high-copy plasmid

Product Specifications

Item number : TD435-CQ-5/50/100/200 (5/50/100/200 times)
Processing capacity : 5–15 mL overnight bacterial culture (OD ≈ 3)
Plasmid size : Up to 20 kb
Elution volume : ≥50 μl (100 μl recommended)

Save and Operate

P1+RNase A : Store at 2–8°C
P2/P4/Washing Solution/Elution Solution : Store at room temperature
Operating temperature : Room temperature (15–30°C)
Operation Time : About 16 minutes

Plasmid DNA Rapid Mini‑to‑Mid‑Scale Extraction Kit – Frequently Asked Questions

1: What are the main features and typical applications of this kit?

This kit is specifically designed for the rapid extraction of plasmid DNA from 5–15 mL of bacterial culture, with the entire procedure completed in just 16 minutes and requiring no phenol/chloroform/ethanol precipitation. Its proprietary colorimetric assay (P1 red, P2, P4) provides intuitive monitoring of the lysis‑neutralization process. The purified product contains endotoxin levels ≤0.5 EU/μg, making it suitable for sensitive applications such as restriction digestion, transformation, PCR, in vitro transcription, sequencing, and transfection.

• Suitable for mid-scale extraction of high-copy plasmids, with a single‑run yield of approximately 100 μg.

• For research use only (RUO).

2: How does the colorimetric reaction help assess the lysis and neutralization states?

After adding P1 (red), the bacterial pellet becomes uniformly rose‑red; upon addition of P2 and gentle inversion, the solution turns a clear, translucent pale pink, indicating complete lysis. Following the addition of P4 and further inversion until the red color completely disappears and the solution turns yellow, a large amount of white flocculent precipitate forms, signifying complete neutralization.

• If red or pink areas remain after neutralization, it indicates insufficient neutralization; continue inverting until the solution turns uniformly yellow.

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

3: Why is it unnecessary to use phenol, chloroform, and ethanol precipitation? What about safety?

• 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; finally, the purified plasmid DNA is eluted with DNase‑ and RNase‑free 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, shortening the procedure time, 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 for transfection?

The purified product exhibits exceptionally high purity: Abs260/280 ≥ 1.8, Abs260/230 ≥ 2.0, endotoxin content ≤ 0.5 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 DNase- and RNase-free water and to ensure that the eluate is free of endotoxin contamination.

• For lower endotoxin levels (e.g., for primary cell transfection), you can combine with an endotoxin removal step or use a dedicated endotoxin‑removal kit.

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

We recommend processing 5–15 mL of overnight‑cultured bacterial culture (OD600 ≈ 3). The typical yield is approximately 100 μg (for high‑copy plasmids). Actual yields may vary depending on factors such as plasmid copy number and the growth state of the host strain.

• If the OD value of the bacterial suspension is too low (<2), the volume may be appropriately increased, but the total bacterial 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 you 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, as this can generate excessive foam and reduce 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 product’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 8–10 times—do not vortex—and incubate for no more than 5 minutes; ③ Immediately after adding P4, invert the tube until the solution turns uniformly yellow and a large amount of white flocculent precipitate forms; ④ After centrifugation, carefully transfer the supernatant, taking care to avoid aspirating any precipitate; ⑤ Before use, add anhydrous ethanol to the wash buffer as instructed and mark the mixture accordingly; ⑥ Prior to elution, spin the sample empty for 1 minute to remove residual ethanol.

• If the OD of the bacterial suspension exceeds 3, the amounts of P1, P2, and P4 can be increased proportionally.

• Preheating the elution buffer to 55–65°C can enhance elution efficiency.

8: Supports both centrifugal and negative-pressure operation modes—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 ideal for high‑throughput processing—when used with a vacuum manifold—and can handle multiple samples simultaneously at a faster pace.

• During vacuum operation, connect the purification column to a 25 mL funnel and ensure a tight seal; apply vacuum 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 DNase/RNase‑free water to 55–65°C, apply it to the purification column matrix, let it stand at room temperature for 5 minutes, and then elute by centrifugation. Preheating can improve elution efficiency by approximately 20–30%.

• If a high‑concentration plasmid is required, 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, and the purified water must be free of DNase and RNase.

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

Take TD435-CQ-100 (100 reactions) as an example: RNase A solution, 2 × 600 μl (4°C); P1 (red), 2 × 60 ml (4°C); P2, 2 × 60 ml (room temperature); P4, 2 × 60 ml (room temperature); Endo‑Free plasmid DNA wash buffer, 2 × 30 ml (room temperature; add ethanol for first use); plasmid DNA elution buffer, 10 ml (room temperature); No. 3 PN purification columns, 2 × 50 units (room temperature); 25 ml funnels, 100 units (room temperature); 2 ml collection tubes, 2 × 50 units (room temperature).

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

• For each bottle of Endo-Free Plasmid DNA Wash Buffer (30 ml), add 120 ml of anhydrous ethanol, mix thoroughly, and mark the bottle to avoid adding it again.

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