Catalog#:
NB-19900
Price/Unit:
$131.00
Unit:
50 preps
For the rapid purification of DNA from various enzymatic reaction mixes.
- Purifies DNA ranging in size from 100 bp – 15000 bp
- Fast and efficient spin-column format
- High DNA recovery
- For use with all types of enzymatic reactions including restriction enzyme digests and Klenow reactions
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BSI's Enzymatic Reaction Clean-Up Kit allows for the rapid purification of DNA from various enzymatic reaction mixes. This kit is able to purify DNA from different enzymatic reactions including restriction enzyme digests, Klenow reactions, alkaline phosphatase reactions, labeling reactions and ligations.
Purification is based on spin column chromatography using BSI's proprietary resin as the separation matrix. Briefly, the enzymatic reaction mixture is first mixed with the provided Binding Solution, and the DNA is then bound to BSI's column (BIND). Under these conditions only the DNA will bind to BSI's resin while any enzymes, reaction by-products and other contaminants are removed in the flowthrough. The bound DNA is then washed to remove any remaining impurities (WASH). Lastly, the purified PCR product is eluted into 30 - 50 µL of the provided Elution Buffer or water (ELUTE). Please see the procedure flowchart to the right.
BSI's resin binds DNA under high salt concentrations and releases the bound DNA under low salt and slightly alkali conditions. The purified DNA can be used in a number of downstream applications including restriction enzyme digestions and ligations.
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FEATURES AND BENEFITS
- Fast and easy processing - Rapid spin-column format allows for the processing of multiple samples in 15 minutes
- Purification from all types of enzymatic reactions - Purify DNA from different enzymatic reactions including restriction enzyme digests, Klenow reactions, alkaline phosphatase reactions, labelling reactions and ligations
- High recovery - Typical purification yield is over 90% of input amount
- Recovered DNA is suitable for downstream applications - Purified DNA is fully compatible with restriction digestions, ligations, sequencing and PCR
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APPLICATIONS
- Restriction digestions
- Ligations
- Sequencing
- PCR
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| Figure 1. Removal of Restriction Enzymes. BSI’s Enzymatic Reaction Clean-Up kit was used to remove the restriction enzyme Hind III from a digestion mixture. Two micrograms of a plasmid containing internal Hind III sites was digested in a 50 µL reaction containing 5 units of Hind III. The restriction digestion was either stopped by adding EDTA or by purification using BSI’s Enzymatic Reaction Clean-up Kit. In all cases, the DNA was incubated at 37°C for 1 hour after the specified treatment. Ten microliters of the digestion or the 50 µL purified DNA were resolved on a 1X TAE, 0.9% agarose gel. Lane 1 contains the control digestion that was immediately stopped by the addition of EDTA. Lane 2 contains control digested DNA. Both lanes 3 and 4 contain digestion reaction that was immediately purified using BSI’s kit. No digestion occurred in lanes 3 and 4, and thus BSI’s Enzymatic Reaction Clean-Up Kit is able to successfully remove Hind III from a reaction mixture. Lane M is BSI’s MidRanger 1kb DNA Ladder. |
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| Figure 2: Removal of DNase I. BSI’s Enzymatic Reaction Clean-Up Kit was used to remove DNase I from a 50 µL reaction. Lanes 1 to 7 of the gel contain DNA that was incubated with increasing amounts of DNase I for 1 hour, and thus act as control lanes for the digestion. Lanes 1 to 7 correspond to 1U, 0.5 U, 0.1 U, 0.05 U, 0.01 U, 0.005 U and 0.0001 U of DNase respectively. Lane 8 is a 50 µL digestion of DNA with 0.5 U of DNase I that was immediately purified after spiking using BSI’s kit. By comparing lane 8 with the other control lanes, it can be determined that BSI’s Enzymatic Reaction Clean-Up Kit can remove DNase I to <0.0001 U, as virtually no digestion occurred. Ten microliters of the digestion or the 50 µL purified DNA were resolved on a 1X TAE, 0.9% agarose gel. |
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| Time to Complete 10 Purifications |
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| Enzymatic Reaction Clean-Up Kit Contents |
| 1. |
Binding Solution |
| 2. |
Wash Solution |
| 3. |
Elution Buffer |
| 4. |
Micro Spin Columns |
| 5. |
Collection Tubes |
| 6. |
Elution Tubes |
| 7. |
Product Insert |
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Long-Term Storage:
All solutions should be kept tightly sealed and stored at room temperature. These 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.