21,00 € – 225,00 €
The EXTRACTME DNA CLEAN-UP & GEL-OUT KIT is designed for a rapid and efficient purification of DNA fragments after enzymatic reactions and directly from agarose gels (standard and low-melting point agarose gels run in either a TAE or TBE buffer). It efficiently removes nucleases, enzyme inhibitors, detergents, restriction enzymes, polymerases, divalent ions, agarose, ethidium bromide and other contaminants. The purified DNA can be used in common downstream applications. The kit enables the purification of DNA fragments from 50 bp to 30 kb, as well as plasmid and genomic DNA. However purification of fragments smaller than 100 bp and larger than 10 kb will result in decreased recovery rates. The purification protocol and buffer formulations were optimized for high yields and purity of DNA. The product is intended for research use only.
- Additional information
- Product information
- Protocol & Manual
- Technical Tips
- Quality & Safety
10 extractions, 250 extractions, 50 extractions
DNA purification procedure utilizes spin minicolumns with membranes which efficiently and selectively bind nucleic acids. In the first step of the clean-up protocol CB Buffer is added to a DNA sample. It causes proteins to degrade and enables DNA binding to the column membrane while in the gel-out protocol DNA fragments is excised from an agarose gel and incubated in GB Buffer, which enables gel fragment solubilization and protein degradation. As an added convenience, the binding buffers contain a color indicator, which facilitates easy monitoring of the solution’s pH for optimal DNA binding. The two-step washing stage efficiently removes impurities and enzyme inhibitors. Purified DNA is eluted with the use either a low ionic strength buffer (Elution Buffer) or water (pH 7.0–9.0) and can be used directly in all downstream applications such as PCR, qPCR, Southern blotting, DNA sequencing, enzymatic restriction, ligation and so forth or stored until ready to use.
CLEAN-UP: up to 100ul of a DNA sample
GEL-OUT: agarose fragment of up to 300 mg containing DNA
Depending on DNA fragment length (in the range of 100 bp – 10 kb):
DNA FRAGMENT LENGTH
100 bp – 10 kb
DNA fragments in the 50–100 bp and 10–30 kbp range can also be purified, as can genomic and plasmid DNA, however the efficiency will be decreased.
Approx. 25 μg DNA
5–10 min for clean-up procedure
16–20 min for gel-out procedure
A260/A280 ratio = 1.7 – 1.9
RECOMMENDATIONS AND IMPORTANT NOTES
GB Buffer contains an indicator, which enables pH monitoring. Yellow indicates that the solution’s pH is lower than 7.5, which guarantees optimal DNA binding with the membrane. When the pH is higher than 7.5, the solution turns pink. It usually happens, for example, when the running buffer for electrophoresis has been used several times or was incorrectly prepared. In such cases, it is essential to add 10 μl of 3 M sodium acetate (pH 5.2). It will lower the pH, enabling the solution to bind efficiently to the minicolumn membrane.
Protocol & Manual
EXTRACTME DNA CLEAN-UP & GEL-OUT KIT (EM26) - Manual
EXTRACTME DNA GEL-OUT KIT – TROUBLESHOOTING
Quality & Safety
The quality of each production batch (LOT) of the EXTRACTME DNA CLEAN-UP & GEL-OUT KIT is tested using standard QC procedures. Purified DNA concentration and quality are evaluated by gel electrophoresis and spectrophotometer.