diff --git a/base/BUILD.gn b/base/BUILD.gn
index 00f206dd70336..7136d40fab12e 100644
--- a/base/BUILD.gn
+++ b/base/BUILD.gn
@@ -1974,10 +1974,6 @@ test("base_unittests") {
       ":base_java_unittest_support",
       "//base/android/jni_generator:jni_generator_tests",
     ]
-
-    # TODO(brettw) I think this should not be here, we should not be using
-    # isolate files.
-    isolate_file = "base_unittests.isolate"
   }
 
   if (is_ios) {
diff --git a/cc/BUILD.gn b/cc/BUILD.gn
index fd1e9c8df7f2d..829f4e3592c87 100644
--- a/cc/BUILD.gn
+++ b/cc/BUILD.gn
@@ -994,10 +994,6 @@ test("cc_perftests") {
     "//ui/gl",
     "//ui/gl:test_support",
   ]
-
-  if (is_android) {
-    isolate_file = "cc_perftests.isolate"
-  }
 }
 # When adding support for isolates, please have a look at run-time dependencies
 # in the cc_unittests_run target in cc_tests.gyp.
diff --git a/chrome/test/BUILD.gn b/chrome/test/BUILD.gn
index 5d6654926ca74..72929f866a48c 100644
--- a/chrome/test/BUILD.gn
+++ b/chrome/test/BUILD.gn
@@ -1700,7 +1700,6 @@ test("unit_tests") {
     android_manifest =
         "//chrome/test/android/unit_tests_apk/AndroidManifest.xml"
     enable_multidex = true
-    isolate_file = "../unit_tests.isolate"
 
     # Some android targets still depend on --gc-sections to link.
     # TODO: remove --gc-sections for Debug builds (crbug.com/159847).
diff --git a/components/BUILD.gn b/components/BUILD.gn
index b2b39d0bd7de2..09cf2b6672292 100644
--- a/components/BUILD.gn
+++ b/components/BUILD.gn
@@ -208,8 +208,6 @@ test("components_unittests") {
   }
 
   if (is_android) {
-    isolate_file = "components_unittests.isolate"
-
     deps += [
       "//components/gcm_driver/instance_id:test_support",
       "//components/gcm_driver/instance_id/android:instance_id_driver_java",
@@ -428,7 +426,6 @@ if (!is_ios) {
         "//testing/android/native_test:native_test_support",
       ]
       use_default_launcher = false
-      isolate_file = "components_browsertests.isolate"
       android_manifest = "${target_gen_dir}/components_browsertests_manifest/AndroidManifest.xml"
       deps += [
         ":components_browsertests_assets",
diff --git a/content/test/BUILD.gn b/content/test/BUILD.gn
index ba81cefd09ae5..b2dde284540e8 100644
--- a/content/test/BUILD.gn
+++ b/content/test/BUILD.gn
@@ -537,8 +537,6 @@ test("content_browsertests") {
     deps -= [ "//device/battery" ]
     android_manifest =
         "${target_gen_dir}/content_browsertests_manifest/AndroidManifest.xml"
-    isolate_file = "//content/content_browsertests.isolate"
-
     use_default_launcher = false
   }
 
diff --git a/media/BUILD.gn b/media/BUILD.gn
index cc5d331cef677..238c8dc534014 100644
--- a/media/BUILD.gn
+++ b/media/BUILD.gn
@@ -590,7 +590,6 @@ test("media_unittests") {
   data_deps = []
 
   if (is_android) {
-    isolate_file = "media_unittests.isolate"
     sources -= [
       "filters/decrypting_audio_decoder_unittest.cc",
       "filters/decrypting_video_decoder_unittest.cc",
@@ -706,7 +705,6 @@ test("media_perftests") {
   ]
   if (is_android) {
     deps += [ "//ui/gl" ]
-    isolate_file = "media_perftests.isolate"
   }
   if (media_use_ffmpeg) {
     deps += [ "//third_party/ffmpeg" ]  # Direct dependency required to inherit config.
diff --git a/media/midi/BUILD.gn b/media/midi/BUILD.gn
index 3d0e60f178598..dc5f0a2e9e8a8 100644
--- a/media/midi/BUILD.gn
+++ b/media/midi/BUILD.gn
@@ -178,10 +178,7 @@ test("midi_unittests") {
 
   if (is_android) {
     deps += [ ":midi_java" ]
-    isolate_file = "midi_unittests.isolate"
-  }
-
-  if (!is_android) {
+  } else {
     sources += usb_midi_sources
   }
 
diff --git a/net/BUILD.gn b/net/BUILD.gn
index 2debac60f1051..9c77a457ac0a8 100644
--- a/net/BUILD.gn
+++ b/net/BUILD.gn
@@ -1627,7 +1627,6 @@ test("net_unittests") {
     set_sources_assignment_filter([])
     sources += [ "base/address_tracker_linux_unittest.cc" ]
     set_sources_assignment_filter(sources_assignment_filter)
-    isolate_file = "net_unittests.isolate"
     shard_timeout = 300
   }
 
diff --git a/sql/BUILD.gn b/sql/BUILD.gn
index 00b7ff68eef3d..fb1bd2f470953 100644
--- a/sql/BUILD.gn
+++ b/sql/BUILD.gn
@@ -114,28 +114,4 @@ test("sql_unittests") {
     "//testing/gtest",
     "//third_party/sqlite",
   ]
-
-  if (is_android) {
-    isolate_file = "sql_unittests.isolate"
-  }
-
-  # TODO(GYP)
-  #['OS == "android"', {
-  #  'dependencies': [
-  #    '../testing/android/native_test.gyp:native_test_native_code',
-  #  ],
-  #}],
-}
-
-if (is_android) {
-  #TODO(GYP)
-  #'target_name': 'sql_unittests_apk',
-  #'type': 'none',
-  #'dependencies': [
-  #  'sql_unittests',
-  #],
-  #'variables': {
-  #  'test_suite_name': 'sql_unittests',
-  #},
-  #'includes': [ '../build/apk_test.gypi' ],
 }
diff --git a/sync/BUILD.gn b/sync/BUILD.gn
index 844a528408829..4e0ffebe31c65 100644
--- a/sync/BUILD.gn
+++ b/sync/BUILD.gn
@@ -724,10 +724,6 @@ test("sync_unit_tests") {
     "//third_party/zlib",
   ]
 
-  if (is_android) {
-    isolate_file = "sync_unit_tests.isolate"
-  }
-
   if (is_chromeos) {
     # Required by get_session_name_unittest.cc on Chrome OS.
     deps += [ "//chromeos" ]
diff --git a/testing/test.gni b/testing/test.gni
index b699fcf53a123..0a6b98674a8ae 100644
--- a/testing/test.gni
+++ b/testing/test.gni
@@ -79,18 +79,11 @@ template("test") {
     }
 
     _test_runner_target = "${_output_name}__test_runner_script"
-    _wrapper_script_vars = [
-      "isolate_file",
-      "shard_timeout",
-    ]
+    _wrapper_script_vars = [ "shard_timeout" ]
     _gen_isolate_vars = [
       "allow_odd_runtime_deps",
       "ignore_all_data_deps",
     ]
-
-    # TODO(agrieve): Delete all manually passed-in .isolate files now that they
-    #     are unused. http://crbug.com/589318
-    assert(!defined(invoker.isolate_file) || invoker.isolate_file != "")  # Mark used.
     _generate_device_isolate =
         !defined(invoker.ignore_all_data_deps) || !invoker.ignore_all_data_deps
 
diff --git a/ui/base/BUILD.gn b/ui/base/BUILD.gn
index ed57bffcd7d75..8fe9f68bc87c1 100644
--- a/ui/base/BUILD.gn
+++ b/ui/base/BUILD.gn
@@ -880,7 +880,6 @@ test("ui_base_unittests") {
       "//chrome:resources",
       "//ui/android:ui_java",
     ]
-    isolate_file = "ui_base_tests.isolate"
   }
 
   if (use_pango) {