Endotoxin Removal Kits- For DNA
For research use only and NOT intended for in vitro diagnostics.
Endotoxin Removal Kits- For DNA
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Supporting Data
Kit Specifications
|
|
Maximum DNA Input
|
25 μg
|
Final Endotoxin Level
|
≤ 0.1 EU/µg DNA
|
Size of DNA Purified
|
Up to 13,000 bp
|
Maximum DNA Volume Input |
100 μL
|
Average Recovery
|
> 90%
|
Time to Complete 10 Purifications |
20 minutes
|
Storage Conditions and Product Stability
All solutions should be kept tightly sealed and stored at room temperature. This kit is stable for 2 years after the date of shipment.
Component | Cat. 22700 (25 preps) | Cat. 52200 (10 preps) | Cat. 21900 (4 preps) |
---|---|---|---|
Buffer SK | 15 mL | 60 mL | 60 mL |
Wash Solution H | 18 mL | 18 mL | 18 mL |
Elution Buffer I | 6 mL | 12 mL | 12 mL |
Endotoxin Removal Solution | 1.5 mL | 1.5 mL | 1.5 mL |
Precipitation Solution | - | 1.5 mL | 1.5 mL |
Spin Columns (assembled with Collection Tubes) | - | 10 | 4 |
Spin Columns | 25 | - | - |
Collection Tubes | 25 | - | - |
Elution Tubes (1.7 mL) | 25 | - | - |
Elution Tubes (15 mL) | - | 10 | - |
Elution Tubes (50 mL) | - | - | 4 |
Product Insert | 1 | 1 | 1 |
Documentation
FAQs
Mini, Midi, Maxi
Poor DNA recovery could be due to one or a combination of the following factors:
- DNA did not bind properly to the column.
Ensure that the Buffer SK does not contain any precipitates. Warm and mix gently if necessary.
- The appropriate amount of ethanol was not added to Wash Solution H.
Wash Solution H has been specifically designed to contain the appropriate amount of components. Ensure that Wash Solution H is prepared using the correct amount of ethanol.
- Incomplete removal of Wash Solution H.
Ensure that the column is spun for specified time minutes after the wash step, in order to completely dry the column. Traces of ethanol may remain in the eluted sample otherwise, and interfere with subsequent enzymatic reactions.
- The appropriate amount of Buffer SK was not added.
Ensure that 5 volumes of Buffer SK is added for every 1 volume of DNA processed. The DNA volume must not exceed the specified volume for each kit.
If the DNA does not perform well in downstream applications, it may be due to one or more of the following:
- DNA was not washed with the provided Wash Solution H.
Ensure that the column is washed with the appropriate amount of Wash Solution H. This solution has been specifically designed to remove all traces of ethanol and endotoxin.
- Proper Elution Buffer was not used.
The provided Elution Buffer I has been optimized for endotoxin-free recoveries. If endotoxin-free water is used for the elution, ensure that the pH is between 7 and 8.
- A different Elution Buffer was used.
The provided Elution Buffer I has been optimized for endotoxin-free recoveries. The endotoxin-free properties of the eluted DNA will be compromised if another elution buffer is used. If a different elution buffer other than the one provided is used, the buffer should also be checked for any components that may interfere with the application. Common components that are known to interfere are high salts (including EDTA), detergents, and other denaturants. Check the compatibility of your elution buffer with the intended use.
Endotoxin levels in the eluted DNA could become slightly higher than 0.1 EU/μg DNA due to the following:
- A different Elution Buffer was used.
The provided Elution Buffer I has been optimized for endotoxin-free recoveries. The endotoxin-free properties of the eluted DNA will be compromised if another elution buffer is used. If a different elution buffer other than the one provided is used, the buffer should also be checked for endotoxin levels.
- The endotoxin levels of the input were extremely high.
If the initial input DNA had extremely high endotoxin levels, the levels may not be completely reduced to 0.1 EU/μg of DNA or less. In this case, the eluted DNA could be applied to a second column and the procedure repeated in order to further reduce the endotoxin levels.