> For the complete documentation index, see [llms.txt](https://knowledge.illumina.com/llms.txt). Markdown versions of documentation pages are available by appending `.md` to page URLs; this page is available as [Markdown](https://knowledge.illumina.com/instrumentation/novaseq-6000/instrumentation-novaseq-6000-troubleshooting-list/000005499.md).

# Troubleshooting NovaSeq 6000 Reagent Chiller outside acceptable range warning

The NovaSeq 6000 reagent chiller remains at room temperature when the instrument is idle. At the start of a sequencing run, the instrument will begin cooling the reagent chiller compartment (RCA) on the instrument and will target a temperature of 7-8°C and maintain that temperature for the duration of the run. Issues with maintaining the appropriate reagent chiller temperature can cause run quality issues.

Issues during the run, if present, will often present the following **warning message**:

21-06-08 13:28:28.818 -05 0208 WRN: The chiller returned a temperature of 23.0289993286133 at the beginning of sequencing, which is outside the acceptable range of \[4, 14] -- SequenceCommand.cs, CheckChillerTemperature, 2729

**Troubleshooting steps**:

* Check if the ambient temperature is significantly higher than expected.
  * Maintain a lab temperature of 19°C to 25°C (22°C ± 3°C). For further information, see [NovaSeq Sequencing System Site Prep Guide](https://support.illumina.com/downloads/novaseq-site-prep-guide-1000000019360.html).
* Collect Run Cycle logs: C:\ProgramData\\\*\Illumina\NovaSeq\NovaSeqTemp\[run folder]\Logs (zipped)
  * After collecting the logs, contact [Illumina Technical Support](https://www.illumina.com/company/contact-us.html#/united-states/technical-support) to determine the next steps.

\
\
\ <br>

|                                                                                                                                                                                                                                                                                                                                                                        |
| :--------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------: |
| *For any feedback or questions regarding this article (Illumina Knowledge Article #5499), contact Illumina Technical Support* [*techsupport@illumina.com*](mailto:techsupport@illumina.com?subject=Question%2FFeedback%20Regarding%20Illumina%20Knowledge%20Article%20#000005499%20-%20Instrumentation%20\&body=Dear%20Illumina%20Technical%20Support,%0D%0A%0D%0A)*.* |


---

# Agent Instructions
This documentation is published with GitBook. GitBook is the documentation platform designed so that both humans and AI agents can read, navigate, and reason over technical content effectively. Learn more at gitbook.com.

## Querying This Documentation
If you need additional information that is not directly available in this page, you can query the documentation dynamically by asking a question.

Perform an HTTP GET request on the current page URL with the `ask` query parameter:

```
GET https://knowledge.illumina.com/instrumentation/novaseq-6000/instrumentation-novaseq-6000-troubleshooting-list/000005499.md?ask=<question>
```

The question should be specific, self-contained, and written in natural language.
The response will contain a direct answer to the question and relevant excerpts and sources from the documentation.

Use this mechanism when the answer is not explicitly present in the current page, you need clarification or additional context, or you want to retrieve related documentation sections.