diff --git a/src/coreclr/nativeaot/BuildIntegration/Microsoft.NETCore.Native.Publish.targets b/src/coreclr/nativeaot/BuildIntegration/Microsoft.NETCore.Native.Publish.targets
index 32eefb307eb0fd..b3219708a8cd13 100644
--- a/src/coreclr/nativeaot/BuildIntegration/Microsoft.NETCore.Native.Publish.targets
+++ b/src/coreclr/nativeaot/BuildIntegration/Microsoft.NETCore.Native.Publish.targets
@@ -55,8 +55,6 @@
 
     <Error Condition="'$(PublishTrimmed)' == 'false'" Text="PublishTrimmed is implied by native compilation and cannot be disabled." />
 
-    <Error Condition="'$(PublishAot)' == 'true' and '$(PublishSingleFile)' == 'true'" Text="PublishAot and PublishSingleFile cannot be specified at the same time." />
-
     <!-- Fail with descriptive error message for common unsupported cases. -->
     <Error Condition="'$(DisableUnsupportedError)' != 'true' and '$(OS)' == 'Windows_NT' and '$(_targetOS)' != 'win'"
       Text="Cross-OS native compilation is not supported." />