Auto-Mag® DTR

Auto-Mag® DTR utilizes AMD’s proprietary magnetic bead technology and is a high-performance reagent designed for the efficient removal of BigDye terminators and sequencing reaction contaminants from Sanger sequencing reactions, ensuring reliable and high-quality DNA sequence reads.

The Auto-Mag® DTR workflow involves selective binding of DNA to paramagnetic beads, followed by the removal of excess nucleotides, primers, and non-target amplicons through washing steps, and concluding with the elution of purified DNA.

Compatible with both manual and fully automated workflows, Auto-Mag® DTR provides a flexible and reliable solution for sequencing reaction cleanup. The protocol is readily adaptable to common liquid handling systems, supporting high-throughput processing.

Key Features

• Efficient Removal of BigDye Terminators
  Effectively removes unincorporated dye terminators and sequencing contaminants
• Improved Sequencing Quality
  Enhances signal clarity and enables longer, high-quality read lengths
• Magnetic Bead-Based Purification
  Streamlined workflow with selective DNA binding, washing, and elution
• Reliable Contaminant Removal
  Eliminates excess nucleotides, primers, salts, and non-target amplicons
• Automation-Ready
  Compatible with common liquid handling systems for high-throughput processing
• Flexible Workflow Integration
  Suitable for both manual and fully automated sequencing cleanup workflows
• Cost-Effective Alternative
  Provides consistent performance without the need for precipitation or column-based methods

Sanger Sequencing Cleanup Workflow

1. Confirm the volume of Sanger sequencing reaction samples.
2. Add 10μl of Auto-Mag® DTR and the corresponding volume of 85% of Ethanol for DNA binding.
3. Separate beads from contaminants.
4. Wash beads/DNA twice with 85% of Ethanol to remove contaminants.
5. Elute purified DNA from beads.
6. Transfer eluate to new vessel and ready for sequencing. 

Data & References 

1. Comparison of Sanger Sequencing Clean-Up Reagents

Fig 1. The 1.4 kb PCR fragments were sequenced using Applied Biosystems’ BigDye Terminator v3.1 Chemistry and cleanup with Auto-Mag® DTR, or other company's sequencing cleanup reagent for comparison.  the DNA samples were analyzed on an Applied Biosystems’ 3730XL.

1. The sequencing chromatogram 1:  the samples purified with Auto-Mag® DTR.
2. The sequencing chromatogram 2: the sample purified with company A sequencing cleanup reagent.
3. The sequencing chromatogram 3: the unpurified samples.

No dye blobs or ski slope effects were observed in sequencing samples purified using Auto-Mag® DTR or Company A’s cleanup reagents, in contrast to unpurified samples.

2. Contiguous read length (CRL) of sequencing sample after cleanup with Auto-Mag® DTR

Fig 2. A 10 µl sequencing reaction sample was purified using 10 µl of Auto-Mag® DTR. The DNA was analyzed on an Applied Biosystems 3730XL sequencer. No dye blobs were observed, and the Contiguous Read Length (CRL) exceeded 1000 bp.