12/10/2022
This bulletin provides information to help calculate the final concentration of formamide in the waste solution of Illumina sequencing runs.
Formamide concentration in the final waste solution of each run varies depending on the instrument and the length of the run. To determine the concentration of formamide, measure the final waste volume and perform the appropriate calculations as explained in the following table.
| Instrument | Version | Formamide Reagent |
Position | Formamide Reagent Fill Volume (mL) |
Formamide Waste Concentration Calculation |
|---|---|---|---|---|---|
| iSeq 100* | i1 | Unlabeled | N/A | 2.00 | 2.0 mL/144.5 mL = 1.4% |
| MiniSeq | v1 | Denature Reagent | 9 | 5.47 | (Formamide reagent)/ (Total waste solution) = % *Use fill volume if remaining formamide is also manually pour or automatically purged into waste solution. |
| MiSeq | v2 | LDR | 8 | 2.48 | |
| v3 | LDR | 8 | 2.48 | ||
| NextSeq 500/550 | v2.5 - Mid Output | Denature Reagent | 6 | 16.89 | |
| v2.5 - High Output | Denature Reagent | 6 | 27.09 | ||
| HiSeq 1000/1500/ 2000/2500 |
High Output - v3 | AT2 | 15 | 5.50 | |
| High Output - v4 | FDR | 15 | 7.05 | ||
| Rapid v2 | FDR | 15 | 5.04 | ||
| HiSeq 3000/4000 | v1 | HDR | 15 | 10.56 | |
| HiSeq X | v2 | PDR | 15 | 10.56 | 10.56 mL/1.3 L = 0.8% |
| NextSeq 1000/2000 | P1, P2 | Unlabeled | N/A | 1.74 | With optional purge. Kits less than 600 cycles 1.74mL/170mL = 1.02%** 600 cycle kit 1.74mL/300.8mL =0.58%** |
| P3 | Unlabeled | N/A | 1.74 | With optional purge 1.74mL/187.8mL = 0.93%** |
|
| NovaSeq | SP, S1, & S2 | Denature Reagent | 30 | 9.00 | 10% |
| S4 | Denature Reagent | 30 | 22.00 | 10% |
*iSeq 100 reagent cartridge is solid waste as it is not intended to be opened.
**Without purge formamide concentration can range from 0.35 - 1.5% depending on run cycles
Example: A HiSeq X v2 run delivers about 10.56 ml of formamide reagent to the waste container. The estimated volume of total waste is about 1.3 L. The percent of formamide in the final waste solution is approximately 0.8%.
Safely dispose formamide with appropriate Personal Protective Equipment as usual. Regulations for the proper disposal of formamide and other hazardous components in the waste vary by region. Contact your Environmental Health and Safety (EH&S) office for the definition of hazardous materials and how to dispose of containers and unused contents in accordance with the governmental safety standards for your region. See the Safety Data Sheets (SDS) on the Illumina support page for more information about chemicals found in Illumina reagents.