Auto-Mag® DTR

Auto-Mag® DTR is designed to effectively and reliably remove BigDye™ terminator from Sanger sequencing reactions to reduce background signal for higher QV scores and reliable long readings of DNA sequences.

• No protocol changes against major competitor.
• Rapid and reliable clean-up of Sanger sequencing reactions
• Longer Phred 20 read lengths averaging over 800 bases with a low retest rate
• Reduce BigDye usage, due to increased average signal strength
• Performed manually or fully automated for high-throughput dye-terminator removal
• Up to 95% significant cost savings compared to any similar products.

The process consists of a selective binding of DNA to the Auto-Mag® DTR particles, followed with washing off nucleotides, primers and non-targeted amplicons, and finally elution of pure DNA to be directly used in subsequent applications. Auto-Mag® DTR is compatible with common sequencing chemistries. The high sensitivity of Auto-Mag® DTR binding capacity allows the use of lower concentrations of BigDye® chemistry and achieves longer continuous read lengths. Auto-Mag® DTR can be processed in both 96- and 384-well formats and is an excellent choice for both manual and fully automated purification of sequencing products.

Auto-Mag® DTR Manual Workflow for Sanger sequencing cleanup

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.

There were no dye blobs or ski slope effect observed at the purified sequencing samples by using Auto-Mag® DTR, or other company A sequencing cleanup reagent.

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

Fig 2. The 10ul of Sequencing reaction sample was purified by using 10ul of Auto-Mag® DTR. The DNA was analyzed on an Applied Biosystems’ 3730XL. There is no dye blods and Contiguous Read Length (CRL) is over 1000bp.