Sequencing Reaction Cleanup

Montage SEQ₉₆ Cleanup Kit and MultiScreen SEQ₃₈₄ Filter Plate

Montage SEQ₉₆ Sequencing Reaction Cleanup Kit

Membrane-based size-exclusion filter plates deliver quality sequencing results

   • 10-minute vacuum-based protocol
   • Compatible with a variety of templates
   • Optimized for use with BigDye chemistries
   • 96- and 384-well formats available
   • Automation compatible

Fast, Vacuum-based Protocol

SEQ filter plates from Millipore incorporate patented size-exclusion technology to yield highly purified sequencing reaction products. In addition to eliminating centrifugation steps, the plates do not require filtrate collection or column packing.

The vacuum-driven protocol is designed for use with automated DNA sequencers, including ABI PRISM® and Amersham Biosciences MegaBACE® systems.

Improved Consistency

Size-exclusion purification eliminates variability in sequencing reaction cleanup. Since the membrane-based protocol does not require alcohol precipitation, there is no risk of salts or ethanol affecting final sequencing results. In addition to high pass rates, highly reproducible results are obtained with PCR and plasmid templates prepared by a variety of methods.

Protocol

1.	Introduce samples into wells.
2.	Filter using Montage vacuum manifold for 6 minutes or until wells are empty. DNA is retained on the membrane surface while salts and dye terminators are filtered to waste.
3.	Wash, resuspend, and recover DNA.

Performance

Montage SEQ₉₆ Sequencing Reaction Cleanup Kit with PCR Template

A 657 bp PCR product (10 fmole) was purified using Montage PCR_µ96 filter plate and sequenced in a 1/8 x BigDye Terminator v1.1 reaction. After purification using the Montage SEQ₉₆ Sequencing Reaction Cleanup Kit, the samples were analyzed on an ABI PRISM 3700 DNA sequencer. Using this approach, the first base after the primer is readily detectable.

Faster Protocols, Improved Results
Method	Process Time	Typical Phred 20 Values	Average Pass Rate
Montage SEQ96 kit	10 minutes	680	>97
Brand Q 96-well gel-filtration-based dye terminator removal system*	55 minutes	630	94
*Manufacturer's recommended protocol

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