Magnetic beads sequencing method of dye Removal Kit | DyeClean Kit | Terminator Removal Kit

DyeClean Kit

For research only

*Preps calculated according to the sample volume of 10 μL

Introduction:

 DyeClean Kit utilizes a unique paramagnetic bead-based purification system for Sanger post sequencing reaction access dye terminator clean-up. The protocol mainly consists of binding, washing and elution steps, and can be performed directly in the thermal cycling plate and requires no centrifugation or filtration.

Process Overview:

1.       Preparation the sample.

2.       Bind sequencing extension products to magnetic beads.

3.       Then separate on magnet plate.

4.       Wash beads to remove unincorporated dyes, nucleotides, salts, and other contaminants.

5.       Elute DNA using aqueous buffer.

6.       Transfer to new plate.

Materials Supplied in the DyeClean Kit:

- Store at 4°C upon arrival, for up to 6 months.

- Mix PuriMag DyeClean well before using.

- DO NOT FREEZE.

 magnetic rack:

For 96 well format: 96R Magnet Plate [# MRack08, http://www.purimagbead.com]

For single tube format: 12 Hole Magnet Rack [# MRack02, http://www.purimagbead.com]

Reagents:

-80% Ethanol made from non-denatured ethanol

n  Make 25 mL of 80% ethanol per 96 well platex

n  Ethanol is used for precipitation in the DyeClean protocol, so it is critical that the 80% ethanol has to be made fresh. To avoid loss of product, make only as much as will be used in 1-3 days and store in a tightly capped container.

-Elution Buffer: Reagent grade water or 0.1mM EDTA (pH 8.0)

The optimal elution buffer will vary depending on dye chemistry and reaction conditions. See individual protocols for details.