Endotoxin-free plasmid mini-prep kit
21-minute rapid extraction, ultrapure and endotoxin-free
One-click copy of information
Instruction manual- Product Description
- Composition
- Precautions
-
Product Overview
This product employs the classic alkaline lysis method, combined with a column-based silica‑membrane technology and an improved endotoxin‑removal solution, to isolate high‑quality plasmid DNA from 1–5 mL of overnight bacterial cultures, effectively eliminating residual endotoxins. The procedure is simple and rapid, allowing complete plasmid extraction in as little as 21 minutes.
The residual endotoxin level in the extracted plasmid DNA is extremely low (＜0.1 EU/μg), making it suitable for direct use in cell transfection, animal injection, restriction digestion, PCR, sequencing, and other experiments.
Storage conditions
Store Solution L of this product at 2–8°C; store the other components at room temperature. If RNase A has been completely added to Solution P1 and thoroughly mixed, Solution P1 should be stored at 2–8°C for a shelf life of 6 months.
Pre-experiment preparation
1. Before first use, add all of the RNase A to Solution P1, mix thoroughly, and then label the tube.
2. Prepare 1.5/2 mL centrifuge tubes, 80% ethanol, and other necessary materials.
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 the cell pellet in the same 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 suspension until it is completely resuspended.
Note: If the bacterial cells are not fully resuspended and dispersed, this will affect the subsequent alkali lysis step, thereby reducing the yield of plasmid DNA. Solution L can enhance plasmid extraction efficiency; please 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 it remains cloudy, it may indicate an excessive cell density or incomplete lysis; in such cases, reduce the amount of bacterial cells.
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 immediately to prevent localized precipitation.
5. Transfer the supernatant to a new 2 mL centrifuge tube, add an endotoxin‑removal solution equal to 0.5× the volume 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 adsorption 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 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 to 100 μL of ddH₂O. H 2 O Place the sample 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.
-
Product Composition
Component Net weight/50 T Item number Solution P1 13 mL 400-201A Solution P2 13 mL 400-201B Solution P3 13 mL 400-201C Except endotoxin solution 20 mL 400-201D Protein removal solution 28 mL 400-201E RNase A 260 μL 400-201F Solution L 260 μL 400-201G dd H 2 O 10 mL 400-201H Adsorption Column & Collection Tube 50 sets 400-201I -
Precautions
1. Solution P2 may precipitate at low temperatures. If precipitation occurs, dissolve the precipitate in a 37°C water bath 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 volatilization and changes in pH.
Keywords
GOONIE
Previous
Product Recommendations
Content is being updated.
Submit your message
If you are interested in our products, please provide your email address, and we will contact you as soon as possible. Thank you!
Goonie Biotechnology (Guangzhou) Co., Ltd.
Room 321–332, Building G5, No. 39 Ruihe Road, Huangpu District, Guangzhou
Customer Service QR Code
Friendly Links
