17,00 € – 230,00 €The EXTRACTME DNA CLEAN-UP KIT is designed for a rapid and efficient purification of DNA fragments after enzymatic reactions. It efficiently removes nucleases, primers, enzyme inhibitors, detergents, restriction enzymes, polymerases, divalent ions, salts, mineral oil, etc. Purified DNA may be used in common downstream applications. Primers from PCR reactions are eliminated quantitatively while small DNA fragments are still bound and purified with high recovery. The kit enables purification of DNA fragments from 50 bp to 20 kbp, as well as plasmid and genomic DNA. The purification protocol and buffer formulations were optimized for high yields and purity of DNA. The product is intended for research use only.
DNA purification procedure utilizes spin minicolumns with membranes which efficiently and selectively bind nucleic acids. First, CB Buffer is added to DNA sample. It causes proteins to degrade and enables DNA to bind with the column’s membrane. The binding buffer contains a color indicator, that facilitates an 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 using either a low ionic strength buffer (Elution Buffer) or water (pH 7.0-9.0) and may be used directly in all downstream applications such as PCR, qPCR, DNA sequencing, enzymatic restriction, DNA ligation, etc. or stored until ready to use.
up to 200 μl of DNA sample
60-99% – depending on DNA fragment length
DNA FRAGMENT LENGTH
50 bp – ~20 kbp
genomic and plasmid DNA, however the efficiency will be decreased
approx. 40 μg DNA
10 min for 6 PCR purifications
A260/280 ratio = 1.7-1.9
RECOMMENDATIONS AND IMPORTANT NOTES
The optimal volume of Elution Buffer used should be chosen in accordance with the amount of DNA in a sample and to final DNA concentration expected. The use of 15-30 μl of Elution Buffer is recommended. It is essential to apply Elution Buffer precisely onto the centre of the membrane. In order to maximize DNA recovery, the following modifications should be applied:
- Heat Elution Buffer up to 70°C, apply onto the column and incubate at room temperature for 5 min.
- Carry out 2 or 3 elution steps with 15-30 μl Elution Buffer. To assure complete DNA recovery from the membrane, use 200 μl Elution Buffer.
Elution Buffer does not contain EDTA, which may interfere with some enzymatic reactions.
CB Buffer contains an indicator, which enables pH monitoring. Yellow indicates that the solution’s pH is lower than 7.0 and guarantees an optimal DNA binding with the membrane. When the pH is higher than 7.0, solution turns pink. It usually happens when the pH of DNA sample considerably differs from the standard parameters of DNA treatment operations (pH>9.0). In this case, 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 with the minicolumn membrane.
Loading Buffer is provided for analysis of purified DNA samples with the use of gel electrophoresis. Loading Buffer contains 3 dyes (bromophenol blue, xylene cyanol and orange G). Loading Buffer is concentrated by a factor of six, thus, in order to obtain the most satisfying results mix 2 μl of Loading Buffer with 10 μl of purified DNA.
EXTRACTME DNA CLEAN-UP KIT (EM07.1) - NEW VERSION - Protocol
EXTRACTME DNA CLEAN-UP KIT (EM07.1) - NEW VERSION - Manual
EXTRACTME DNA CLEAN-UP KIT – TROUBLESHOOTING
The quality of each production batch (LOT) of the EXTRACTME DNA CLEAN-UP KIT is tested with the use of standard QC procedures. Purified DNA concentration and quality are evaluated by gel electrophoresis and spectrophotometer.
EXTRACTME DNA CLEAN-UP KIT (EM07.1) - NEW VERSION - MSDS
EXTRACTME DNA CLEAN-UP KIT (EM07.1) - NEW VERSION - CoA L.736472
EXTRACTME DNA CLEAN-UP KIT (EM07.1) - NEW VERSION - CoA L.756472
EXTRACTME DNA CLEAN-UP KIT (EM07.1) - NEW VERSION - CoA L. 785673
EXTRACTME DNA CLEAN-UP KIT (EM07.1) - NEW VERSION - CoA L. 786273
EXTRACTME DNA CLEAN-UP KIT (EM07.1) - NEW VERSION - CoA L. 816473
EXTRACTME DNA CLEAN-UP KIT (EM07.1) - NEW VERSION - CoA L. 826473
EXTRACTME DNA CLEAN-UP KIT (EM07.1) - NEW VERSION - CoA L. E07706673
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