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DNA Extraction from PAGE Gels
                              Applications for Oligonucleotide Purification




A well tried and established method to purify synthetic oligonucleotides, is the so called “soak and crush
method”. Briefly the method, consist of:

    1. After PAGE electrophoresis is completed, the oligonucelotide bands are visualized by the UV
        shadowing method
    2. Using a scalpel, the oligonucleotide bands are carefully cut from the gel.
    3. The bands are gently crushed and cut in to smaller fragments and placed into a test tube containing
        5mls ( or more, but alewyas enough to cover all the crushed gel)of 0.1 X TE(Tris EDTA)
    4. The crushed gel is left for 3 hours at room temperture(or overnight if convenient; 37 degrees incubation,
        renders higher recovery and is optional.
    5. A 1 cm slurry of DE-52 resin ( this is a positively charge resin, that will retain the polyanionic
        oligonucleotide) is prepared in a 10 cm plastic column with medium or coarse fritted filter
    6. The liquid 0.1X TE,now containing the oligonucleotide is passed through the DE-52 bed
    7. The column is washed 3x with 10mls each of dd water
    8. Use a 1 molar solution of TEAB( triethylammonium bicarbonate buffer) to elute the oligo
    9. Use 3x1 ml of the TEAB buffer
    10. Using a speed vac, evaporate the TEAB buffer and measure your A260 absorbance; oligo is ready for
        your use


Bio-Synthesis.Inc, has been producing synthetic oligonucleotides for over 25 years; not only DNA, but
RNA, other modified oligonucleotides, and provide cross-linking of various types of biomolecule using
our optimized bioconjugation strategies, which have a number of applications on gene expression
inhibition and related anti-sense studies. Also synthetic peptides and peptide antibodies for a number of
biological relevant applications, in the areas of proteomics, epigenetics, immune regulation, post
translational modifications, antisense, gene expression control, RNA interference and more. For more
product information, visit: www.biosyn.com.

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Dna%20 extraction%20from%20page%20gels

  • 1. DNA Extraction from PAGE Gels Applications for Oligonucleotide Purification A well tried and established method to purify synthetic oligonucleotides, is the so called “soak and crush method”. Briefly the method, consist of: 1. After PAGE electrophoresis is completed, the oligonucelotide bands are visualized by the UV shadowing method 2. Using a scalpel, the oligonucleotide bands are carefully cut from the gel. 3. The bands are gently crushed and cut in to smaller fragments and placed into a test tube containing 5mls ( or more, but alewyas enough to cover all the crushed gel)of 0.1 X TE(Tris EDTA) 4. The crushed gel is left for 3 hours at room temperture(or overnight if convenient; 37 degrees incubation, renders higher recovery and is optional. 5. A 1 cm slurry of DE-52 resin ( this is a positively charge resin, that will retain the polyanionic oligonucleotide) is prepared in a 10 cm plastic column with medium or coarse fritted filter 6. The liquid 0.1X TE,now containing the oligonucleotide is passed through the DE-52 bed 7. The column is washed 3x with 10mls each of dd water 8. Use a 1 molar solution of TEAB( triethylammonium bicarbonate buffer) to elute the oligo 9. Use 3x1 ml of the TEAB buffer 10. Using a speed vac, evaporate the TEAB buffer and measure your A260 absorbance; oligo is ready for your use Bio-Synthesis.Inc, has been producing synthetic oligonucleotides for over 25 years; not only DNA, but RNA, other modified oligonucleotides, and provide cross-linking of various types of biomolecule using our optimized bioconjugation strategies, which have a number of applications on gene expression inhibition and related anti-sense studies. Also synthetic peptides and peptide antibodies for a number of biological relevant applications, in the areas of proteomics, epigenetics, immune regulation, post translational modifications, antisense, gene expression control, RNA interference and more. For more product information, visit: www.biosyn.com.