Auto-Mag® DTR

• Magnetic Bead-Based High-Quality, Cost-Efficient Dye Terminator Removal Reagent
• No protocol changes required compared to major competitors
• Rapid and reliable cleanup of Sanger sequencing reactions
• High DNA recovery yields with long sequence reads (over 800 bps)
• Suitable for manual use or adaptable to automated liquid handlers

Auto-Mag® DTR utilizes AMD's proprietary magnetic beads and is a high-performance reagent designed for the removal of BigDye terminators and sequencing reaction contaminants from Sanger sequencing reactions, ensuring reliable and long DNA sequence reads. The Auto-Mag® DTR process involves selective binding of DNA to paramagnetic beads, followed by washing away nucleotides, primers, and non-target amplicons, and concluding with the elution of pure DNA. Auto-Mag® DTR is an excellent choice for both manual and fully automated purification of sequencing reaction products. The protocol is adaptable to common liquid handling workstations.

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.