|Oligo Conjugation to magnetic beads for affinity matrix
Click here for Magnetic Bead Conjugation Specifications
This modification is a post synthesis NHS conjugation to a primary amino group thus an additional modification with an amino group is required. A C6 or C12 amino group can be placed at the 5' or for the 3' end a C3 or C7 amino and for internal positions an amino modified base is used, e.g Amino dT C6.
Oligo Design. A stretch of T's of a length of 15 to 20mer is recommended to be added to the end of the oligo that is conjugated to the magnetic particles to avoid steric hindrance and to ensure optimum hybridization.
1 micromolar scale: ~5 mg magnetic bead conjugated to ~2 nmol oligo. 10mg/mL suspension.
2 micromolar scale: ~10 mg magnetic bead conjugated to ~5 nmol oligo. 10mg/mL suspension.
Gene Link offers a wide variety of modifications that enable oligonucleotides to be conjugated to various ligands including solid surfaces with the appropriate functional groups. The most common conjugation chemistry involves oligos modified with a primary amino group; the amino group can be placed at 5' or 3' end or any desired internal position using an amino C6 base. The amino group reacts with N-hydroxysuccinimide(NHS) functional groups to form a stable amide linkage; the reaction efficiency varies from 50% to 90% depending on the ligand activated NHS group. The requirement for this conjugation is the availability of the desired ligand or solid surface in an activated NHS form for conjugation to the amino modified oligonucleotide.
Material Supplied & Handling
1. The oligo conjugated to magnetic beads are supplied at 10mg/mL.
2. The Oligo-Magnetic bead conjugation varies between 0.3-0.5 nmol amine oligo/mg magnetic bead. Refer to specification sheet supplied with product and label on the tube.
3. Wear appropriate personal protective equipment and clothing including lab coat, safety glasses and gloves.
4. The conjugated oligo-magnetic bead is supplied in High Salt Buffer (0.5 M NaCl, 50 mM Tris-HCl pH 7.5 and 0.02% sodium azide.
5. Sodium Azide Note: Dilute solutions of sodium azide are used in research laboratories as a preservative. This use generally presents no extraordinary dangers to the user, but it should be noted that weak solutions of sodium azide (0.1 to 1.0%) are eye and skin irritants.
6. Refer to the oligo specification sheet for the exact nmol oligo/mg of magnetic bead.
DO NOT CENTRIFUGE. DO NOT FREEZE. Store the supplied oligo-magnetic bead at 4oC.
1. Do not centrifuge the beads as it will collapse and form aggregates that will be difficult to re-suspend.
2. Use magnetic stand for separating the beads from the solution for all applications.
3. The material supplied is in 0.02% sodium azide (High Salt Buffer (0.5 M NaCl, 50 mM Tris-HCl pH 7.5 and 0.02% azide). Handle with caution.
4. Handle with caution. Before initial use perform a few washes in the binding buffer of choice to equilibrate the new binding buffer and removal of azide.
5. For DNA/oligo binding & release. Use High Salt ((0.5 M NaCl, 50 mM TrisHCl pH 7.5) for binding and Low Salt (50 mM Tris-HCl ph 7.5) or sterile water for elution. Elution at 60oC is preferable.