EXTRACTME PLASMID MIDI ENDOTOXIN-FREE kit (EM17)

EXTRACTME PLASMID MIDI ENDOTOXIN-FREE kit (EM17)

143,00 305,00 

Extraction kit - EXTRACTME PLASMID MIDI ENDOTOXIN-FREE kit (EM17) and EXTRACTME PLASMID MIDI kit (EM16) are designed for the efficient purification of high quality plasmid DNA from 50-300 ml of cultured bacterial cells. The kits are based on anion-exchange resins, allowing extraction of ultrapure, transfection-grade pDNA, which is highly suited for use in demanding applications. The isolation protocol and buffer  formulations were optimized for high isolation efficiency and pDNA purity. The product is intended for research use only.

Extraction kit: EM17, EM17-010, EM17-025

Available Options:

PRINCIPLE

The plasmid DNA purification procedure utilizes pre-packed anion-exchange resin columns which efficiently and selectively bind nucleic acids. In the first isolation step, the pDNA is released from bacterial cells by alkaline lysis. Then the lysate is neutralized and all the cell residues along with the proteins and genomic DNA are separated in the centrifugation step. This alkaline lysis method and RNase treatment are used to obtain clear cell lysate with minimal genomic DNA/RNA contaminants. In the next step the lysate is  applied to the purification column with the equilibrated resin and the DNA is bound. The washing stage effectively removes impurities and enzyme inhibitors. A suitable buffer with a high ionic strength allows the elution of the plasmid DNA, which is then concentrated and desalted by precipitation. The purified plasmid DNA may be used directly in all downstream applications such as PCR, qPCR, transfection, microinjection, Southern blotting, DNA sequencing, enzymatic restriction and so forth, or stored until ready to use.

PRODUCT SPECIFICATIONS

SAMPLE MATERIAL

Bacterial broth culture:
50-150 ml (high-copy number plasmids)
100-300 ml (medium- and low-copy number  plasmids)

YIELD

200-600 μg of transfection-grade pDNA from 100 ml of cultured bacterial cells

TIME REQUIRED

  • 70 min for EM16 and 100 min for EM17
  • additional ~60 min for DNA precipitation

DNA PURITY

A260/A280 ratio = 1.7 – 1.9

ENDOTOXIN REMOVAL (EM17)

<0.1 EU/μg verified by LAL

RECOMMENDATIONS AND IMPORTANT NOTES

EXTRACTME PLASMID MIDI ENDOTOXIN-FREE kit – TROUBLESHOOTING

QUALITY CONTROL

The quality of each production batch (LOT) of the EXTRACTME PLASMID MIDI kits are tested using BLIRT’s ISO-certified quality management system. The purified DNA concentration, quality and stability are evaluated by gel electrophoresis and spectrophotometry. In addition, the  functional quality is tested by qPCR, digestion with restriction enzymes and pDNA transfection.

EXTRACTME PLASMID MIDI ENDOTOXIN-FREE kit (EM17) - MSDS