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DyeEx 2.0 Spin Kit

For removal of unincorporated dye terminators from 1–24 sequencing reactions

  • Fast procedure with only two short centrifugation steps
  • Ready-to-use prehydrated gel-filtration material
  • Efficient removal of any dye terminator

DyeEx Kits provide either spin columns or 96-well plates and use gel-filtration technology to remove dye terminators from sequencing reactions. Sequencing reactions are loaded onto the pre-hydrated gel-filtration material. After a short centrifugation step, the reactions are ready to be loaded onto a capillary sequencer. Unincorporated dye terminators are retained in the gel matrix.

Артикул: 63206X4
DyeEx 2.0 Spin Kit (1000)
1000 DyeEx Spin Columns, Collection Tubes (2 ml)
Артикул: 63206
DyeEx 2.0 Spin Kit (250)
250 DyeEx Spin Columns, Collection Tubes (2 ml)
Артикул: 63204
DyeEx 2.0 Spin Kit (50)
50 DyeEx Spin Columns, Collection Tubes (2 ml)

Performance

DyeEx Kits remove any type of dye terminator from 10–20 µl sequencing reactions including BigDye, dRhodamine dye, Rhodamine, DYEnamic ET, and WellRED terminators. After cleanup, the sequencing  reactions can be separated on any capillary sequencer. We recommend the DyeEx 96 Kit using the optimized protocol. 

High-quality sequencing

DyeEx Kits are optimized for fast and convenient dye-terminator removal leading to high-quality sequencing results. In contrast, sequencing-reaction cleanup by ethanol precipitation is very time consuming (see table "Comparison of DyeEx procedure and ethanol precipitation") and inefficient. The failure to remove dye terminators efficiently often leads to the appearance of dye blobs in sequencing data making stretches of sequence unreadable. Using DyeEx Kits for sequencing reaction cleanup ensures that the reactions loaded onto sequencing instruments are free of dye terminators. Without dye blobs, the sequence is easily readable throughout. Signal intensities are high, resulting in long read lengths.

Comparison of DyeEx procedure and ethanol precipitation

Procedure  DyeEx 2.0 Spin Ethanol
precipitation
Time required*
(12 samples, spin format)
10 minutes >45 minutes
Time required*
(Microplate format for 96 samples)
N.A. >60 minutes
Handling Ready-to-use
spin format
Multiple
pipetting steps
Sequence quality ++  +
N.A.: not applicable
* Does not include time required to dry down samples before gel loading (where applicable).
Using the standard protocol.

Principle

The DyeEx dye-terminator removal system is designed for fast and efficient removal of unincorporated dye terminators from sequencing reactions using a simple spin column procedure. The DyeEx procedure uses gel filtration to quickly and efficiently remove unincorporated terminators from sequencing reactions. Removal of dye terminators is important to prevent the unincorporated dye terminators from interfering with analysis of sequencing results. The DyeEx gel-filtration material consists of spheres with uniform pores and separates molecules according to molecular weight. When sequencing reaction mixtures are applied to DyeEx columns, dye terminators diffuse into the pores and are retained in the gel-filtration material, while labeled DNA fragments are excluded and recovered in the flow-through.

Procedure

Dye-terminator removal with DyeEx Kits is fast because the procedure is so simple. A quick centrifugation step removes storage buffer from the columns, the sequencing samples are loaded, and a second centrifugation step removes unincorporated dye terminators. Samples are then ready for direct loading onto a capillary sequencer or can be dried, redissolved, and loaded onto a sequencing gel.

Applications

After cleanup, the sequencing reactions can be separated on any capillary sequencer.

Features
Specifications
Binding capacity 10–20 µl
Elution volume 10–20 µl
For DyeEx Kits: Terminator removal, ABI PRISM 377, 373, 310, 3100, or 3700, MegaBACE 1000, or CEQ 2000 sequencers
Format Tube/96-well plate
Processing Manual
Removal Dye terminator
Sample type: applications Sequencing reactions
Technology Gel filtration