Sample Preparation Products

For the rapid removal of endotoxins from previously purified DNA

Reduce endotoxin levels to 0.1 EU/µg DNA or less Effectively remove endotoxins in as little as 20 minutes Remove endotoxins from up to 40 µg of DNA Rapid spin-column format

BSI’s Endotoxin Removal Kit (Mini) is designed for the rapid removal of endotoxins from up to 40 µg of previously purified DNA. Endotoxins are released during the lysis step of plasmid purification and significantly reduce transfection efficiencies in endotoxin sensitive cell lines. Therefore, the removal of endotoxins from plasmid preparations is often necessary prior to the use of the DNA in downstream applications. With BSI’s Endotoxin Removal Kit (Mini) endotoxin levels are efficiently reduced to 0.1 EU/µg DNA or less.

Purification is based on spin-column chromatography using BSI’s proprietary resin as the separation matrix. Briefly, the DNA sample is first mixed with the provided Binding Solution, the sample is placed into the top reservoir of the column, and Endotoxin Removal Solution is added. The column is then spun in order to bind the DNA (BIND). Under these conditions only the DNA will bind to BSI’s resin while the contaminating endotoxins are removed in the flow-through. The bound DNA is then washed to remove any remaining impurities (WASH). Lastly, the endotoxn-free plasmid DNA is eluted into 50 - 100 µL of the provided Elution Buffer or water (ELUTE). Please see the procedure flowchart to the right.

The Endotoxin Removal Kit (Mini) can be used to purify plasmids up to 13,000 bp in size, with average recoveries of greater than 90%. The purified DNA is of the highest quality and can be used in a number of downstream applications including sequencing, cloning and transfection.

FEATURES AND BENEFITS

• Fast and easy processing - Rapid spin-column format allows for the processing of multiple samples in 20 minutes
• Endotoxin-free plasmid DNA - Using BSI ’s Endotoxin Removal Kit (Mini), endotoxin levels are reduced to less than or equal to 0.1 EU/µg plasmid DNA
• High recovery of input DNA - Average DNA recovery is > 90% of the input amount
• Recovered DNA is suitable for downstream applications - Purified DNA is fully compatible with sequencing, cloning and transfections

APPLICATIONS
Purified DNA is of the highest quality and can be used in a number of downstream applications including:

• Sequencing
• Cloning
• Transfections

Figure 1. Efficiently Remove Endotoxins. An input of 11 µg of a plasmid in a 50 µL volume with an endotoxin level of over 8 EU/µg was subjected to clean-up by BSI ’s Endotoxin Removal Kit. One microliter of the input and the 50 µL eluted DNA was diluted 1000-fold or 10 fold, respectively for a 100 µL end-point Limulus Amebocyte Lysate test for endotoxin level. The endotoxin levels of the plasmid DNA sample were reduced from over 8 EU/µg to less than or equal to 0.1 EU/µg of plasmid DNA using BSI’s Endotoxin Removal Kit (Mini), with high DNA recovery also (see Figure 2).

Figure 2. High DNA Recovery. An input of 11 µg of a plasmid in a 50 µL volume with an endotoxin level of over 8 EU/µg was subjected to clean-up by BSI’s Endotoxin Removal Kit in triplicate. The 50 µL DNA elutions was quantified by spectrophotometry. There is very little loss of DNA associated with using BSI’s Endotoxin Removal Kit (Mini). Recoveries of plasmid DNA are greater than 90% of the input amount when using this kit with efficient endotoxin removal (see Figure 1.)

Endotoxin Removal Kit (Mini) - For DNA Contents
1.	Binding Solution
2.	Wash Solution
3.	Elution Buffer
4.	Endotoxin Removal Solution
5.	Micro Spin Columns
6.	Collection Tubes
7.	Elution Tubes
8.	Product Insert

Long-Term Storage: All solutions should be kept tightly sealed and stored at room temperature. All the reagents should remain stable for at least 1 year in their unopened containers.

Product Usage: For laboratory research use only, not for diagnostic use! Not for use in humans.

Reference/Citations: