Plasmid Mini Prep Kit
Simple and fast operation—extraction completed in 35 minutes.
One-click copy of information
Instruction manual- Product Description
- Composition
- Precautions
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Product Overview
This product employs the classic alkaline lysis method and utilizes a column-based silica‑membrane technology, enabling the high‑quality extraction of plasmid DNA from 1–5 mL of overnight bacterial cultures while effectively removing contaminants such as proteins and polysaccharides. The procedure is simple and rapid, with plasmid isolation completed in as little as 21 minutes.
The extracted plasmid DNA exhibits high yield and purity, making it suitable for direct use in restriction digestion, ligation, transformation, PCR, sequencing, and other experiments.
Storage conditions
The solution L of this product should be stored at 2–8°C, while the other components may be stored at room temperature. If RNase A has been fully added to solution P1 and thoroughly mixed, solution P1 must be stored at 2–8°C and is stable for 6 months.
Pre-experiment preparation
1. Before first use, add all of the RNase A to Solution P1, mix thoroughly, and then label the container.
2. Prepare 1.5/2 mL centrifuge tubes, 80% ethanol, isopropanol, and other necessary reagents.
Instructions for Use
1. Pipette 1–5 mL of the overnight culture into a 2-mL centrifuge tube, and centrifuge at 14,000 × g for 1 minute to pellet the cells. (If the culture volume is large, perform multiple centrifugations to collect all the cells in a single tube.) Carefully aspirate and discard the supernatant, ensuring that all liquid is removed.
2. Sequentially add 250 μL of Solution P1 (already containing RNase A) and 5 μL of Solution L, and vortex the bacterial pellet until it is completely resuspended.
Note: If the bacterial cells are not completely resuspended and dispersed, this will affect the subsequent alkaline lysis step, thereby reducing the yield of plasmid DNA. Solution L can enhance plasmid extraction efficiency; therefore, add 5 μL of Solution L to Solution P1 in the centrifuge tube first to avoid loss.
3. Add 250 μL of Solution P2, gently invert the tube 8 times, and let it stand at room temperature for 3 minutes (do not exceed 5 minutes).
Note: Do not vigorously vortex during this step; mix gently to avoid residual genomic DNA. The lysis time should not exceed 5 minutes to prevent plasmid degradation. At this point, the bacterial suspension should become clear and viscous. If the suspension remains cloudy, it may indicate an excessive amount of cells or incomplete lysis; in such cases, reduce the cell density.
4. Add 250 μL of Solution P3, immediately invert the tube 15 times; a white flocculent precipitate should form. Centrifuge at 14,000 × g for 10 minutes.
Note: After adding solution P3, mix thoroughly immediately to prevent localized precipitation.
5. Transfer the supernatant to a new 2 mL centrifuge tube, add isopropanol at a volume equal to 0.5 times that of the supernatant, and invert the tube 15 times to ensure thorough mixing.
6. Transfer the mixture from Step 5 into the adsorption column (which has already been placed in the collection tube), centrifuge at 14,000 × g for 30 seconds, and discard the supernatant.
Note: The maximum loading capacity of the spin column is 750 μL. If the volume of the mixture exceeds this limit, it must be loaded in multiple aliquots.
7. Repeat step 6 until the entire mixture has been loaded onto the column.
8. Add 500 μL of protein‑removal solution to the adsorption column, let it stand at room temperature for 30 seconds, centrifuge at 14,000 × g for 30 seconds, and discard the supernatant.
9. Add 700 μL of 80% ethanol to the adsorption column, let stand at room temperature for 30 seconds, centrifuge at 14,000 × g for 30 seconds, and discard the supernatant.
10. Place the adsorption column back into the collection tube and centrifuge at 14,000 × g for 1 minute.
11. Place the adsorption column into a new 1.5 mL centrifuge tube, and add 30 μL–100 μL of dd. H 2 O Place the membrane at the center of the adsorption column, let it stand at room temperature for 1 minute, centrifuge at 14,000 × g for 1 minute, and discard the adsorption column.
12. The extracted plasmid DNA was stored at −20°C.
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Product Composition
Component Net content/100 T Item number Solution P1 26 mL 400-200A Solution P2 26 mL 400-200B Solution P3 26 mL 400-200C Protein removal solution 53 mL 400-200D RNase A 520 μL 400-200E Solution L 520 μL 400-200F dd H 2 O 12 mL 400-200G Adsorption Column & Collection Tube 100 sets 400-200H -
Precautions
1. Solution P2 may precipitate at low temperatures. If precipitation occurs, dissolve the precipitate by incubating at 37°C until the solution becomes clear before use.
2. After use, each component should be promptly capped to prevent prolonged exposure to air, which may lead to alcohol evaporation and changes in pH.
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Goonie Biotechnology (Guangzhou) Co., Ltd.
Room 321–332, Building G5, No. 39 Ruihe Road, Huangpu District, Guangzhou
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