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Automated Sequencing

Post-Reaction Clean-Up

For removal of unincorporated dyes, excess salts and other interfering components from sequencing reactions.

SigmaSpin™ Post-Reaction Clean-Up Columns

SigmaSpin™ Post-Reaction Clean-Up Columns are ideal for lower throughput applications, such as clean-up of probe labeling reactions or small numbers of sequencing reactions.

These columns can accept sample volumes up to 100 µl. Each column comes with a collection tube to collect the DNA during centrifugation.

SigmaSpin™ 96-Well Post-Reaction Clean-Up Plates

SigmaSpin™ 96-Well Post-Reaction Clean-Up plates provide a fast, simple, and highly efficient method for removing unincorporated dyes, excess salts, and other interfering reaction components (Fig. 1). Each plate is packed with a pre-hydrated size-exclusion resin, equilibrated with molecular biology grade water, and supplied in our unique plate design with long drip directors to minimize contamination between samples. The plate design also includes a snap-cap bottom seal and a foil seal top to ensure that the resin remains hydrated. SigmaSpin™ has been tested in high-throughput genome centers and core facilities with ABI Prism® 3700, 3100, 310 and 377. Each well can accept sample volumes up to 20 µl.

Ideal for removing

  • Dye-terminator nucleotides and primers from sequencing reactions
  • Radiolabeled nucleotides, primers, and fluorescent dyes from nucleic acid probe labeling reactions

SigmaSpin™ Post-Reaction Clean-Up 96-well plates come ready for immediate use.

Features and Benefits

  • Validated with most automated sequencers and dye terminator chemistries (Fig. 2)
  • Pre-qualified size-exclusion resin guarantees optimum performance
  • Unique drip directors prevent cross-contamination between samples during collection
  • Plates are sealed to eliminate leakage or drying during shipping or storage
  • Suitable for use with multi-channel pipettes and automated workstations

Comparison of Sequencing reaction clean-up methods

Panel A: Sequencing reactions were precipitated with 70% ethanol and placed on ice for thirty minutes. DNA pellets were dried and resuspended in TE solution prior to electrophoresis.

Panel B: Sequencing reactions were subjected to post-reaction clean-up with SigmaSpin™ Post-Reaction Clean-Up 96-Well Plates, according to recommended protocol.

Figure 1. Single stranded M13MP18 plasmid was sequenced with a -21M13 forward sequencing primer using ABI BigDye™ Terminator chemistry. Sequencing reactions were resolved on an ABI Prism® 377 XL instrument with a 48 cm gel cassette containing 4.5% AutoPAGE™ Plus acrylamide at 2.88kV for 7 hrs.

SigmaSpin Removes Interfering Reaction Components

Figure 2. Sequencing reaction was purified using a SigmaSpin 96-well plate. pFLAG™ MAC plasmid (Sigma, E5644) was sequenced using BigDye™ Terminator v 3.0 chemistry (20 µl total reaction volume; BigDye premix diluted 1:1 with SeqSaver™ (Sigma, S3938)). Data was generated on an ABI Prism® 3700 DNA Analyzer with POP-6™ polymer and 10X CE Buffer (Sigma, B4930). Phred 20 > 600.

Ordering Information

Product Product Description Quantity
S5059 SigmaSpin™ Post-Reaction Clean-Up Columns (with collection tubes) 70/pkg