Reagent stability and thawing guidelines for the NovaSeq X/X Plus

Background: 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.

Note: All kit components are shipped at room temperature. When the reagents are received, promptly store components at the indicated temperature for proper performance.

Storage requirements

Quantity	Kit Component	Storage Temperature	Notes
1	Reagent cartridge	-25°C to -15°C
1	Lyo insert	-25°C to -15°C	Avoid stacking lyo inserts when storing
1	Custom primer buffer	-25°C to -15°C
1	Pre-load buffer	-25°C to -15°C
1	Flow cell	2°C to 8°C
1	Buffer cartridge	Room temperature
1	Library tube strip	Room temperature

Thawing Guidelines

Reagent Cartridge There are two validated methods for thawing a NovaSeq X/X Plus reagent cartridge.

1. Room Temperature Water Bath.

2. Refrigerator.

Method	Minimum time to complete thaw	Do not exceed	Storage of thawed cartridge
Room Temperature Water Bath (15-30°C)	4 hours	24 hours	72 hours at 2-8°C
Refrigerator (2-8°C)	48 hours	N/A	72 hours at 2-8°C

Note 1: Do not insert the library tube strip or lyo insert into the reagent cartridge while thawing. Make sure to inspect position #30 to verify a complete thaw. Dry thoroughly before placing in the instrument. Note 2: If the reagent cartridge cannot be loaded into the instrument within 24 hours, store at 2°C to 8°C for up to 72 hours or return to -25°C to -15°C storage for up to 7 days. After thawing, do not refreeze more than one time.

Lyo Insert

1. Remove the Lyo Insert from -25°C to -15°C storage.

2. Thaw at room temperature for 10 minutes. Leave Lyo insert in package until ready to use.

3. Thaw Lyo Insert separately. Do not attempt to thaw lyo insert inside of the frozen reagent cartridge.

4. If lyo insert cannot be loaded within 24 hours, return to -25°C to -15°C storage.

Pre-load and Custom Primer Buffers

1. Remove the pre-load and custom primer buffers insert from -25°C to -15°C storage.

2. Thaw at room temperature for 10 minutes and invert 5 times.

3. If pre-load and custom primer buffers cannot be loaded within 8 hours, return to -25°C to -15°C storage.

Flow Cell 1. Remove flow cell from 2°C to 8°C storage. 2. Set the sealed flow cell package aside for 10-15 minutes to allow the flow cell to reach room temperature. 3. Leave flow cell in package until ready to load flow cell into instrument. Use the flow cell within 2 hours of removing it from 2°C to 8°C storage. 4. If flow cell cannot be used within 2 hours, return to 2°C to 8°C storage and use within 24 hours.

For further information, see the NovaSeq X Plus Product Documentation.

For any feedback or questions regarding this article (Illumina Knowledge Article #7561), contact Illumina Technical Support [e-mail].