Auto-Mag® X-Pure Size Select NGS

Auto-Mag® X-Pure Size Select NGS is based on paramagnetic bead technology, and designed for PCR amplicons cleaning up, DNA fragments or RNA purification, and sheared DNA size selection and cleanup in the library construction process for next generation sequencing (NGS).

• Designed for “bottle swap” with no protocol change against major competitor.
• High recovery of PCR amplicons or dsDNA fragments >100bp 
• Single, or double-sided size selection capability with uniform fragments size distribution for NGS
• Rapid and reliable clean-up total RNA, including miRNA, siRNA, and aRNA
• Flexibility of starting sample volumes with the ability to scale and consistent results
• Compatible with manual and automated processes
• Significant cost savings

For PCR products purification, the purification protocols provide the procedures for PCR purification in single tube or 96/384 well format. Auto-Mag® X-Pure Size Select NGS reagent utilizes an optimized buffer to selectively bind DNA fragments >100 bp to the beads. Excess primers, nucleotides, salts, and enzymes can be removed using a simple washing procedure and PCR products are eluted with low salt elution buffer or water.

In the DNA size selection, size selection is required to produce a uniform distribution of fragments around an average size. The protocols of DNA size selection describe procedures of DNA size selection by adjusting volume ratio of Auto-Mag® X-Pure Size Select NGS reagent to sheared DNA sample and the desired size DNA fragment are selected and recovered by a simple binding, washing and elution steps.

Auto-Mag® X-Pure Size Select NGS is RNase free reagent and suitable for RNA cleanup and concentration. The highly purified RNA can be used directly for downstream applications, such as PCR and RT-PCR reactions, RNase protection assays, Transfection for RNAi experiments, Antisense RNA (aRNA) amplification, Probes for microarray or microarray, cDNA synthesis and labeling, as well as RNA-Seq library preparations, etc.

Auto-Mag® X-Pure Size Select NGS is suitable for both manual and fully automated processing and the highly purified DNA can be used directly for downstream applications.

Product name change notice: To intuitively clarify the characteristics and functions of this product, the name of Auto-Mag® X-Pure Select  is changed to Auto-Mag® X-Pure Size Select NGS. There are no changes in the characteristics, quality and usage of the product.

Auto-Mag® X-Pure Size Select Workflow

Left Side Size Selection

1. Bind ds-DNA to magnetic beads.
2. Separate beads from contaminants.
3. Wash magnetic beads with 85% ethanol to remove contaminants.
4. Elute DNA from magnetic beads.
5. Transfer purified DNA to new storage vessels.

 Double Side Size Selection 

1. Confirm the sample volume.
2. Add corresponding volume of Auto-Mag® X-Pure Size Select to the sample for first binding.
3. Separate the beads and transfer supernatant to a new vessels.
4. Add corresponding volume Auto-Mag® X-Pure Size Select to the supernatant for second binding.
5. Separate beads from contaminants.
6. Wash magnetic beads with 85% ethanol to remove contaminants.
7. Elute DNA from magnetic beads.
8. Transfer to new vessels.

Data & References

DNA size selection and Cleanup

Fig 1: DNA size selection was performed using 20 µl of GeneRuler Ultra Low Range DNA Ladder and 100 bp DNA Ladder mix at different ratios of Auto-Mag® X-Pure Size Select NGS volume to DNA sample volume (1.8x to 0.4x). Selected DNA fragments were eluted in 20 µl and analyzed on an Agilent TapeStation 2200.

Comparison of Double-Sided Size Selection

Fig 2. Comparison of electropherograms of  the double-sided size selection on sheared gDNA at 0.7x/0.2x ratio set using Auto-Mag® X-Pure Size Select NGS and a similar kit from Company A following the manufacturer’s recommended protocol. DNA was eluted in 25 µL and analyzed on an Agilent TapeStation® 2200.

Total RNA Clean-Up

Fig 3. The 20μl of total RNA at 300 ng/μl  were cleaned up with Auto-Mag® X-Pure Size Select NGS following recommended protocols. The purified RNA was eluted in 50μl and analyzed on Agilent’s TapeStation® 2200. Recovery rates ranged between 90-95% respectively