Consumables used in Illumina DNA PCR Free workflow

Is there a document describing everything in the kit? Refer to the “Kit Contents” section of the Reference Guide.

Which user-supplied consumables, plastics, and benchtop accessories are needed? Refer to the Reference Guide’s Consumables and Equipment section.

Which pipette tips are recommended? Filter DNase/RNase free low-binding tips.

Can strip tubes be used instead of PCR plates? Strip tubes will work for reactions provided that the thermal cycler can fit them adequately and that the correct magnet is utilized. An example of a magnet that works with strip tubes is the DynaMag-96 Side Magnet (Cat. # 12331D) from Thermo Fisher Scientific.

What are the consequences of using a different type of plate for reactions in thermal cycler and Hybex protocols?

• The thermal cycler protocol uses PCR plates: PCR plates different than recommended will need to be evaluated for several factors, such as volume held in wells and type of magnet needed. The protocol is effective when using 96-well hard-shell plates from BioRad (HSP9601) and DynaMag-96 Side Skirted Magnet (Cat. # 12027). Different magnet/plate combinations may necessitate more careful supernatant removal if magnet concentrates beads at the bottom of the well.

• The Hybex protocol uses MIDI plates: MIDI plates other than recommended must be evaluated for several factors, such as volume held in wells and type of magnet needed. The protocol is effective when using Amgen MIDI plates (Cat. # AB-0859) and Magnetic Stand-96 (Cat. # 10027) are used. Due to MIDI plate design, pipetting steps are substituted with shaking steps, and incubation steps duration and temperatures are adjusted to take into account heat distribution, due to material differences (thickness of the plastic and mass) compared to PCR plates.

Can other kinds of plate seals be used instead of Microseal B adhesive seals?

These are the only seals that have been tested for this protocol by Illumina for thermal steps; however, for libraries storage, Illumina recommends Microseal F foil seals.

For any feedback or questions regarding this article (Illumina Knowledge Article #3448), contact Illumina Technical Support [e-mail].