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, E 5644) was sequenced using BigDye™ Terminator v 3.0 chemistry (20 µl total reaction volume; BigDye premix diluted 1:1 with SeqSaver™ [Sigma, S 3938]). Data was generated on an ABI Prism® 3700 DNA Analyzer with POP-6™ polymer and 10X CE Buffer (Sigma, B 4930). Phred 20 > 600.
Ordering Information
Product Product Description Quantity
S5059 SigmaSpin™ Post-Reaction Clean-Up Columns
(with collection tubes) 		70/pkg
S4309 SigmaSpin™ Post-Reaction Clean-Up Plates* 2 each
S4434 SigmaSpin™ Post-Reaction Clean-Up Plates* 10 each
S4559 SigmaSpin™ Post-Reaction Clean-Up Plates* 50 each
P4736 48-well Wash Plate 50 each
Z37,212-9 96-Well V-bottom Collection Plates 50 each

*Wash and collection plates included in 2- and 10-each package sizes

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