The I-Blue MINI Plasmid Kit was designed for rapid isolation of plasmid DNA from 1-7 mL of cultured bacterial cells. I-Blue Lysis Buffer (an optional color indicator) is included with the kit in order to prevent common handling errors, ensuring efficient cell lysis and neutralization. A modified alkaline lysis method and RNase treatment are used to obtain clear cell lysate with minimal genomic DNA and RNA contaminants. Typical yields are 20-35 µg for high-copy number plasmid or 3-10 µg for low-copy number plasmid from 4 mL of cultured bacterial cells. DNA phenol extraction or alcohol precipitation is not required and the entire procedure can be completed within 15 minutes. The purified plasmid DNA is ready for use in restriction enzyme digestion, ligation, PCR, and sequencing reactions.

Advantages

• Sample: 1-7 mL of cultured bacterial cells
• Yield: Up to 50 µg of pure plasmid DNA
• Format: Plasmid spin column
• Operation Time: Within 15 minutes
• Elution Volume: 30-100 µL
• Kit Storage: Dry at room temperature (15-25°C) for up to 1 year; PD1 and RNase A mixture should be stored at 2-8°C for up to 6 months

Quality Control
The quality of the I-Blue MINI Plasmid Kit is tested on a lot-to-lot basis by isolating plasmid DNA from a 4 mL overnight E. coli (DH5α) culture containing plasmid pBluescript (A600 > 2 U/mL). Following the purification process, a yield of more than 20 µg is obtained and the A260/A280 ratio is between 1.8-2.0. The purified plasmid DNA (1 µg) is used in EcoRI digestion, and analyzed by electrophoresis.

Kit Components

*For IB47171 and IB47172 add provided RNase A to PD1 Buffer then mix by shaking for a few seconds. Check the box on the bottle. PD1 and RNase A mixture should be stored at 2-8°C for up to 6 months. For IB47170 samples, RNase A was already added to PD1.

**If precipitates have formed in PD2 Buffer, warm the buffer in a 37°C water bath, followed by gentle shaking to dissolve.

***Add absolute ethanol (see the bottle label for volume) to Wash Buffer then mix by shaking for a few seconds. Check the box on the bottle. Be sure and close the bottle tightly after each use to avoid ethanol evaporation.

The Fast Ion Plasmid Maxi Kit (Endotoxin Free) uses pre-packed anion exchange resin columns to purify plasmid DNA from 100-250 ml of bacterial cultures. Modified alkaline lysis method (1) and RNase treatment are used for obtaining clear cell lysate with minimal genomic DNA and RNA contaminants. Once the plasmid DNA has been bound to the column, the contaminants can be washed off using the wash buffer. Finally, the purified plasmid DNA is eluted by a high salt buffer and precipitated with isopropanol for desalting. The entire procedure can be completed in 120 minutes and the obtained high purity plasmid DNA is suitable for transfection, sequencing reactions, PCR and in-vitro transcription.

The 96-Well Genomic DNA Kits are designed for high-throughput purification of total DNA (including genomic, mitochondrial and viral DNA) from whole blood and a variety of animal tissues or cells. This method uses Proteinase K and a chaotropic salt to lyse cells and degrade proteins. DNA in the chaotropic salt is bound by the glass fiber matrix of each well. Once any contaminants have been removed, the purified DNA is eluted by a low salt elution buffer or water. The entire procedure can be completed in 1 hour without phenol extraction or alcohol precipitation. These kits can be used for manual filtration or with robotic handling systems, and purified DNA with approximately 20-30kb is suitable for PCR or other enzymatic reactions.

The Genomic DNA Mini Kit for Plants provide a quick and easy method for purifying total DNA (including genomic DNA, mitochondrial and chloroplast DNA) from plant tissue. Samples are disrupted by both grinding in liquid nitrogen and lysis buffer incubation. The lysate is treated with RNase A to degrade RNA and then filtered to remove cell debris and salt precipitates. In the presence of the binding buffer, coupled with chaotropic salt, genomic DNA in the lysate binds to the glass fiber matrix of the spin column. Contaminants are removed using a Wash Buffer (containing ethanol) and the purified genomic DNA is eluted by a low salt Elution Buffer or TE. The procedure does not require DNA phenol extraction or alcohol precipitation, and can be completed in less than 1 hour. The purified genomic DNA is ready for use in PCR, Real-time PCR, Southern Blotting and RFLP.

The Genomic DNA Mini Kit for Tissue was designed specifically for purifying total DNA (including genomic, mitochondrial and viral DNA) from a variety of animal tissue, paraffin-embedded tissue, buccal swab and amniotic fluid. The provided micropestle can efficiently homogenize tissue samples to shorten the time in the Lysis Step. Proteinase K and chaotropic salt are used to lyse cells and degrade protein, allowing DNA to be easily bound by the glass fiber matrix of the spin column. Once any contaminants have been removed, using a Wash Buffer (containing ethanol), the purified DNA is eluted by a low salt Elution Buffer or TE. The entire procedure can be completed within 1 hour without phenol/chloroform extraction or alcohol precipitation. The expected yield of genomic DNA is up to 50µg and the purified DNA (with approximately 20-30 Kb) is suitable for use in PCR or other enzymatic reactions.

The Genomic DNA Kit for Blood or Cultured Cells provides an efficient method for purifying total DNA (including genomic, mitochondrial and viral DNA) from whole blood, frozen blood, buffy coat, cultured animal/bacterial cells and fungus. Chaotropic salt is used to lyse cells and degrade protein, allowing DNA to bind to the glass fiber matrix of the spin column. Contaminants are removed using a Wash Buffer (containing ethanol) and the purified genomic DNA is eluted by a low salt Elution Buffer or TE. The entire procedure can be completed in 1 hour without phenol/chloroform extraction or alcohol precipitation, with an average DNA yield of 6 µ g from 200 µ l of whole human blood and up to 50µ g of DNA from 200µ l of buffy coat. Purified DNA, with approximately 20-30 Kb, is suitable for use in PCR or other enzymatic reactions.