The sequencing workflow begins with the Polymerase Chain Reaction or PCR. In PCR, primers, DNA polymerases, and deoxynucleotides are used to create thousands of copies of the target fragment. Before Sanger sequencing, a PCR cleanup is extremely important to make sure the unincorporated primers and dNTPs do not interfere with the results and to ensure the accuracy of Sanger sequencing.

Steps for the PCR clean up

PCR cleanup can be achieved with different methods, such as a bead or column-based purification, and ethanol precipitation.

Ethanol precipitation is cost-effective but extremely labor-intensive. Bead or column-based is better. But even in the column-based method, samples have to be transferred in and out of the column, leading to the loss of some samples. Magnetic bead-based purification works best for most applications.

Let’s take a closer look to understand it better.

The HighPrep™ DTR Clean-up System includes a magnetic beads-based reagent for manual and automated Sanger sequencing reaction clean-up for both ABI and MegaBACE sequencing platforms. The system lets DNA selectively bind to the paramagnetic beads, followed by washing off nucleotides, primers, and non-targeted amplicons, and finally elution of pure DNA.

The HighPrep™ DTR Clean-up System allows Sanger sequencing reaction clean-up/dye terminator removal for –

• Sanger sequencing on ABI platforms
• Sanger sequencing on MegaBACE platforms

The benefits of the HighPrep™ DTR Clean-up System include rapid and reliable clean-up of sequencing reactions, high DNA recovery yields, and long sequence readings. The system is flexible and can be used in any format, without the need for centrifugation or filtration. It works for both manual and fully automated purification of sequencing reaction products and it is adaptable to common liquid handling workstations.

The HighPrep™ DTR Clean-up System allows -

• selective binding of 100bp and larger DNA fragments
• efficient removal of Primers, Primer‐dimers, dNTPs, salts, enzymes, and other impurities
• avoiding centrifugation/filtration steps

The HighPrep™ DTR Clean-up System cleanup reagent provides a quick, efficient, and convenient answer to your PCR cleanup needs.

If you are looking for a kit for the normalization of DNA concentration or an anger sequencing reaction clean-up kit, call one of the most trusted RNA extraction kit manufacturers in the USA, MagBio Genomics at (301) 302-0144.