- +1 858 909 0079
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- +1 858 909 0079
- [email protected]
Products
One-Step Single-Stranded DNA Removal Kit
Get the Highest Quality DNA Without the Hassle
Applications
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Presequencing clean-up for PCR products
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Megaprimer PCR for site-directed mutagenesis
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Removal of ssDNA containing a 3′-hydroxyl terminus from nucleic acid mixtures
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Assay for the presence of single-stranded DNA with a 3′-hydroxyl terminus
Features and Advantages
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Efficient one-tube and extraction-free protocol
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Ultrafast: Process 96 samples in less than 30 minutes with <10-second Hands-on Time
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Nuclease recovered at the end of the reaction thereby can be reused.
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Easy separation of the nuclease from the reaction.
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Stability of the immobilized nuclease increases.
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Cost-effective: Eliminates columns, filters, laborious, organic reagents, and minimal plasticware required.
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High-throughput: Compatible with many different automated liquid handling systems.
Workflow
Formulation:
Liquid (Supplied in 20 mM Tris-HCl (pH 7.5), 0.1 mM EDTA, 1 mM DTT and 50 % (v/v) glycerol)
Activity:
1 μl Magnetic Beads will digest 0.5μg of oligonucleotides in 67 mM glycine-KOH (pH 9.5), 6.7 mM MgCl2, 1 mM DTT in 30 minutes at 37 °C.
Reaction Buffer:
10X Reaction Buffer 670 mM glycine-KOH (pH 9.5 at 25 °C), 67 mM MgCl2,10 mM DTT.
Shipping:
Shipped at ambient temperature. Upon receipt, store magnetic nuclease beads at -20°C. Aliquot to avoid repeated freezing and thawing.
PROTOCOL
A. Accessory equipment
Magnetic Rack
Item
Magnetic Rack for centrifuge tube
** Based on sample volume, the user can choose one of the following magnetic Racks
Source
• BcMag™ Rack-2 for holding two individual 1.5 ml centrifuge tubes (Bioclone, Cat. No. MS-01)
• BcMag™ Rack-6 for holding six individual 1.5 ml centrifuge tubes (Bioclone, Cat. No. MS-02)
• BcMag™ Rack-24 for holding twenty-four individual 1.5-2.0 ml centrifuge tubes (Bioclone, Cat. No. MS-03)
• BcMag™ Rack-50 for holding one 50 ml centrifuge tube, one 15 ml centrifuge tube, and four individual 1.5 ml centrifuge tubes (Bioclone, Cat. No. MS-04)
Item
BcMag™ 96-well Plate Magnetic Rack.
Source
• BcMa™ 96-well Plate Magnetic Rack (side-pull) compatible with 96-well PCR plate and 96-well microplate or other compatible Racks (Bioclone, Cat. No. MS-05)
Items
Magnetic Rack for centrifuge tube
** Based on sample volume, the user can choose one of the following magnetic Racks
Source
●
BcMag™ Rack-2 for holding two individual 1.5 ml centrifuge tubes (Bioclone, Cat. No. MS-01)
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BcMag™ Rack-6 for holding six individual 1.5 ml centrifuge tubes (Bioclone, Cat. No. MS-02)
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BcMag™ Rack-24 for holding twenty-four individual 1.5-2.0 ml centrifuge tubes (Bioclone, Cat. No. MS-03)
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BcMag™ Rack-50 for holding one 50 ml centrifuge tube, one 15 ml centrifuge tube, and four individual 1.5 ml centrifuge tubes (Bioclone, Cat. No. MS-04)
BcMag™ 96-well Plate Magnetic Rack
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BcMa™ 96-well Plate Magnetic Rack (side-pull) compatible with 96-well PCR plate and 96-well microplate or other compatible Racks (Bioclone, Cat. No. MS-05)
B. Procedure
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Do not use buffers containing organic solvents.
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Typically, the bead is added directly into any standard buffer at the desired amount of the beads based on the concentration of the ssDNA (1 μl Magnetic Beads will digest 0.5μg of oligonucleotides).
1.
Shake the bottle to resuspend the Magnetic beads until it is homogeneous entirely.
IMPORTANT! It is essential to mix the beads before dispensing. Do not allow the beads to sit for more than 2 minutes before dispensing. Resuspend the magnetic beads every 2 minutes.
2.
Add an appropriate amount of the magnetic beads to a reaction.
3.
Mix the sample with beads for 1-2 minutes by slowly pipetting up and down 20-25 times or Vortex the sample for 2 minutes at 2000 rpm.
4.
Incubate at 37°C with continuous rotation for 30 minutes.
5.
Place the sample plate or tube on the magnetic Rack for 30 seconds or until the solution is clear.
(Option: centrifuge at 3500 rpm for 45 seconds)
6.
Transfer the supernatant to a clean plate /tube while the sample plate remains on the magnetic separation plate. The sample is ready for downstream applications.