What is the final formamide concentration in the waste solution of Illumina sequencing systems?


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
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 ((Volume of formamide reagent)/
(Total waste solution volume) = %

Note: 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
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 P2 Unlabeled N/A 1.74 With optional purge
1.74mL/170mL = 1.02%

Note: Without purge
formamide concentration
can range from 0.8 - 1.5%
depending on run cycles
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. 

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 of formamide with appropriate Personal Protective Equipment. 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.