Sequencing reagent performance depends on 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.
iSeq 100 | MiniSeq | MiSeq | NextSeq 500/550 | NextSeq 1000/2000 | HiSeq 1000/1500/2000/2500 | HiSeq 3000/4000/X | NovaSeq
iSeq 100 Reagent Kits
- If thawed in a room temperature water bath (20 to 25°C) for a minimum of 6 hours, the cartridge is stable up to 18 hours in the water bath
- If thawed in a refrigerator (2°C to 8°C) for a minimum of 36 hours, the cartridge is stable up to 72 hours in the refrigerator
- If thawed in room temperature air (20°C to 25°C) for a minimum of 9 hours, the cartridge is stable up to 18 hours in room temperature air
- iSeq 100 i1 Reagent v2 kits can be thawed at 4°C for 36 hours and are stable at 4°C for up to one week prior to use
MiniSeq Reagent Kits
- If thawed in a 37°C water bath for 35 minutes, it is stable up to 2 hours
- If thawed in a room temperature water bath (19°C to 25°C) for 90 minutes, it is stable up to 24 hours
- If thawed at room temperature air (19°C to 25°C) for 5 hours, it is stable up to 24 hours
- If thawed at 2°C to 8°C for 18 hours, it is stable up to 72 hours
MiSeq v2 and v3 Reagent Kits
- If thawed in a room temperature deionized water bath for 60 minutes, place on ice or set aside at 2°C to 8°C and use within 6 hours
- If thawed overnight at 2° to 8°C, store at 2° to 8°C for up to 1 week
NextSeq 500/550 Reagent Kits
- If thawed in a room temperature deionized water bath for 60 minutes, place on ice or set aside at 2°C to 8°C, and use as quickly as possible or within the same day
- If thawed overnight at 2° to 8°C, store at 2° to 8°C for up to 1 week
NextSeq 1000/2000 Reagent Kits
- Water bath: Thaw in temperature controlled 25°C water bath, with a minimum of 9.5 cm water depth for 6-8 hours for 300 cycle or smaller kits; and 600 cycle kits require 8-10 hours.
- Refrigerator (2° to 8°C): One day prior to anticipated run, remove cartridge from -25°C to -15°C storage. Thaw at room temperature for 6 hours. Transfer to refrigerator 2° to 8°C, and continue to thaw for a minimum of 12 hours for 300 cycle or smaller kits; or continue to thaw for a minimum of 16 hours for 600 cycle kits . The cartridge is stable up to 72 hours in the refrigerator. When thawing at room temperature, cartridge should have air on all sides except the bottom and should not be stacked. In the refrigerator, cartridges may be stacked but should be given ample space on all other sides.
- Room temperature (20°C to 25°C) for a minimum of 9-16 hours for 300 cycle or smaller kits; 12 hours (16 hour maximum) for 600 cycle kit.
HiSeq 1000/1500/2000/2500 Cluster and SBS Kits
TruSeq SR Cluster Kit v3, TruSeq PE Cluster Kit v3, HiSeq SR Cluster Kit v4, and HiSeq PE Cluster Kit v4
- If the cBot reagent plate is thawed in a room temperature water bath for 60 minutes, use within the same day
- If the cBot reagent plate is thawed at 2°C to 8°C overnight, no longer than 16 hours, use the plate within the same day
TruSeq Rapid v1 Cluster Kit and HiSeq Rapid v2 Cluster Kit
- If the cBot reagent plate is thawed in a room temperature water bath for 30 minutes, use within the same day
- If the cBot reagent is plate thawed at 2°C to 8°C overnight, no longer than 16 hours, use the plate within the same day
HiSeq v4 High Output SBS Reagents and HiSeq Rapid v2 SBS Reagents
- If the reagents are thawed in a room temperature deionized water bath for about 90 minutes, use within the same day
- If the reagents are thawed at 2°C to 8°C overnight, use the next day
HiSeq 3000/4000/X Cluster and SBS Kits
HiSeq 3000/4000 SR Cluster Kit, HiSeq 3000/4000 PE Cluster Kit, and HiSeq X Five/Ten Cluster Kit
- Thaw the cBot reagent plate in a room temperature water bath for 60 minutes
- Thaw the EPX1, EPX2, EPX3, and RSP reagents at room temperature for 10 minutes, then set on ice
HiSeq 3000/4000 SBS Reagents and HiSeq X Five/Ten SBS Reagents
- If the reagents are thawed in a room temperature deionized water bath for about 90 minutes, use within the same day
- If the reagents are thawed at 2°C to 8°C overnight, use the next day
NovaSeq Reagent Kits
NovaSeq Cluster and SBS cartridges
- Thaw SP, S1, and S2 cluster cartridges in a room temperature deionized water bath for 2 hours
- Thaw S4 cluster cartridges in a room temperature water bath for 4 hours
- Thaw SP, S1, S2, and S4 SBS cartridges in a room temperature water bath for 4 hours
NovaSeq XP Workflow Reagents
- Thaw DPX1, DPX2 and DPX3 at room temperature for 10 minutes, then set aside on ice