Reagent Stability and Thawing Guidelines for MiSeq Sequencing Kits

Sequencing reagent performance depends on the thawing technique and thawed reagent storage time and conditions. It is important to understand and follow the recommendations for reagent thawing and storage because some reagents contain enzymes that are sensitive to temperature and time. Unless specified below, refreezing thawed reagents can negatively affect enzyme activity and run performance.

Storage requirements:

How to prepare reagent cartridges and flow cells for sequencing:

Kit Type	Thawing Method	Storage Method post-thaw	Use by
v2 cartridge	Room temperature deionized water bath for 60 minutes.	Place on ice or set aside at 2°C to 8°C.	Use within 6 hours.
v3 cartridge	Room temperature deionized water bath for 60-90 minutes.	Place on ice or set aside at 2°C to 8°C.	Use within 6 hours.
v2 or v3 cartridge	Refrigerator (2° to 8°C) overnight.	Store at 2° to 8°C.	Use within 1 week.

Reagent kit:

• Thaw the reagent kit in a room temperature deionized water bath with enough water to submerge the base of the reagent cartridge for 60 minutes for the v2 cartridge and 60-90 minutes for the v3 cartridge. Place on ice or set aside at 2°C to 8°C and use within 6 hours.

• Alternatively, thaw reagents overnight at 2° to 8°C, then store at 2° to 8°C for up to 1 week.

• Thawing the reagent cartridge overnight in the refrigerator allows for more flexibility as the reagents are then able to be stored in the refrigerator for up to one week.

Flow cell:

• Remove the flow cell from the fridge and allow it to come to room temperature for 15 minutes.

• Lightly rinse the flow cell with lab-grade water and dry with a lint-free tissue.

• The flow cell glass can be cleaned with an alcohol wipe (be sure not to get any alcohol on the flow cell gaskets).

  • Dry any excess alcohol with a lint-free tissue.

• Inspect the gaskets to ensure they are clear of debris or other abnormalities.

For any feedback or questions regarding this article (Illumina Knowledge Article #1674), contact Illumina Technical Support [e-mail].