# Performing a spike in of custom oligos into an AmpliSeq for Illumina Panel

AmpliSeq for Illumina spike-in panels are usually formulated at 50X concentration. See the steps below for adding a spike-in panel to an AmpliSeq for Illumina ready-to-use or custom panel.

* Most AmpliSeq panels are supplied at 2X concentration (working concentration), though not all. Consult the relevant reference guide to confirm panel concentration and volume usage information.
* Spike-in oligos are sent at 50X concentration.
* Combine the spike-in panel with the main panel 1:25.
  * Typically, this is adding 5 µl AmpliSeq spike-in panel to one tube of main panel pool; confirm in the relevant reference guide before combining oligos.
* Use combined spike-in + main panel pool following the standard library preparation protocol in the relevant reference guide.

Refer to the reference guide for the main AmpliSeq for Illumina panel for additional information on spike-in panel use.

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| *For any feedback or questions regarding this article (Illumina Knowledge Article #1238), contact Illumina Technical Support* [*techsupport@illumina.com*](mailto:techsupport@illumina.com?subject=Question%2FFeedback%20Regarding%20Illumina%20Knowledge%20Article%20#000001238%20-%20Library%20Preparation%20\&body=Dear%20Illumina%20Technical%20Support,%0D%0A%0D%0A)*.* |


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