Library Prep with the I.DOT Liquid Handler & NEBNext Ultra II DNA Library Prep Kit

Learn how the I.DOT tackles every step of the NEBNext Ultra II DNA Library Kit, from fragmentation to pooling, with minimal user intervention.

Did you know that liquid handling automation can save labs hours of hands on time? Welcome to episode two of Library Prep with the I.DOT Liquid Handler. Here, we talk about the NEBNext Ultra II DNA Library Prep Kit. It requires limited amounts of DNA and generates high quality libraries with high yields.

The I.DOT Liquid Handler is designed to dispense microliter scale volumes precisely and accurately, and it can be used with all of the reagents in this kit (Table 1). Here's how.

Learn more about the I.DOT Non-Contact Dispenser

Table 1. Summary of the components in the NEBNext Ultra II DNA Library Prep Kit.

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You can use the I.DOT to dispense your fragmentation enzyme and then, during end repair, use the I.DOT to dispense the end prep enzyme mix and the end prep reaction buffer. The I.DOT can dispense up to 65% glycerol, so even a highly viscous enzyme can easily be dispensed with this liquid handler.

Then during adaptor ligation, the I.DOT can dispense the adapter for Illumina, the ligation master mix, and the ligation enhancer.

At any point during your library prep that you need to do a clean-up, whether it's post fragmentation, post ligation, at the very end after PCR, you can use the I.DOT to dispense your magnetic beads and NEB offers this kit with sample purification beads.

You can take your library prep to the next level by adding the G.PURE NGS Clean-up Device. The G.PURE is special because it not only can clean your samples, but it will dispense the ethanol and resuspension buffer, as well. The G.PURE also reduces your lab's tip consumption, reducing it by up to five tip boxes per cleanup.

DISPENDIX I.DOT Liquid Handler & G.PURE NGS Clean-Up Device

Add in the fact that you can automate size selection, as well, using your beads, the I.DOT and G.PURE combo can automate up to 80% of your library prep workflow.

Next for PCR, the I.DOT can dispense both the master mix and the primers.

Finally, pool using the I.DOT source plates with variable volumes of library and water.

That's it! No training required and no automation experience necessary. Book a demo today!


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