YALEPIC^® Ultrapure Plasmid DNA Mini Extraction Kit

This kit is routinely recommended for 1-5 mL of overnight culture. For low-copy plasmids, the culture volume can be appropriately increased to <=10 mL, and Buffer PA, PB, and PM2 reagent volumes should be strictly scaled up proportionally to ensure complete lysis, neutralization, and binding steps for improved yield.

Common causes include aged bacteria, overgrowth of culture, incomplete lysis, or insufficient binding/elution time. Recommendations: re-pick a fresh single colony for overnight culture (12-16 hours optimal), strictly control lysis time within 5 minutes, extend binding or elution time. Preheating Elution Buffer to 60 degrees C before use helps improve elution rate. For low-copy plasmids, appropriately increase culture volume and extend binding time.

This kit is pre-mixed with a blue/yellow indicator: the lysis buffer should normally be blue. If it turns yellow-green or yellow, this usually indicates abnormal buffer system pH or excessive sample addition. Reduce the culture volume and ensure all buffers are fresh and free of precipitate (if precipitate appears, dissolve in a 37 degrees C water bath before use). If the mixture does not become fully clear after neutralization, check whether neutralization is sufficient or mix again and centrifuge.

Yes. Gram-positive bacteria require cell wall disruption pre-treatment (e.g., lysozyme treatment). After cell wall disruption, follow this kit’s standard operating protocol (alkaline lysis method + spin column purification) for extraction.

For large plasmids >10 kb, 5-10 mL of culture is recommended, with PA/PB/PM2 Buffer volumes strictly increased proportionally. Binding and elution steps can be appropriately extended in reaction time. Use Elution Buffer preheated to 60 degrees C and avoid vigorous shaking to prevent large plasmid breakage.

This kit is shipped at ambient temperature (10-30 degrees C) and stored at room temperature. If Buffer PB shows precipitate before use, dissolve in a 37 degrees C water bath and mix thoroughly. The unopened kit has a shelf life of 12 months.

The maximum yield can reach 45 microgram of plasmid DNA (high-copy plasmid, 5 mL culture). Both high-copy and low-copy plasmids are applicable. For low-copy plasmids, increase culture volume and scale up reagent amounts to achieve optimal extraction efficiency.

Spin column clogging is typically caused by incomplete lysis or excessive culture volume. Recommendations: use a vortexer for thorough mixing during lysis to avoid residual bacterial clumps. Strictly follow the 1-5 mL recommended culture volume to avoid overloading. Ensure centrifugation force is >=12,000 x g.

The YALEPIC Ultrapure Plasmid DNA Mini Extraction Kit uses imported specialty silica membrane material and an optimized high-compatibility alkaline lysis buffer system, delivering up to 45 microgram yield with A260/A280 purity of 1.8-2.0. Combined with the blue/yellow indicator for visualized operation, the extraction efficiency and purity meet the requirements of downstream applications such as molecular cloning, sequencing, and transfection, with significantly better cost-effectiveness than comparable imported and joint-venture plasmid extraction products.

Eluted plasmid DNA can be stored at -20 degrees C for short-term (within 1 week). For long-term storage, aliquot and store at -80 degrees C to avoid repeated freeze-thaw. Use sterile, nuclease-free ddH2O or Elution Buffer for elution to improve storage stability.

This kit provides RNase A. Before use, add all RNase A to Buffer PA, mix well, aliquot, and store at 2-8 degrees C. Avoid RNase contamination and use RNase-free consumables during operation to further improve extraction quality.