diff --git a/api/src/opentrons/protocol_engine/types.py b/api/src/opentrons/protocol_engine/types.py
index d5b126542d4..ad861051b6c 100644
--- a/api/src/opentrons/protocol_engine/types.py
+++ b/api/src/opentrons/protocol_engine/types.py
@@ -909,3 +909,5 @@ class EnumParameter(RTPBase):
 
 
 RunTimeParameter = Union[IntParameter, FloatParameter, EnumParameter]
+
+RunTimeParameterValues = Dict[str, Union[float, bool, str]]     # update value types as more RTP types are added
\ No newline at end of file
diff --git a/api/src/opentrons/protocol_reader/protocol_reader.py b/api/src/opentrons/protocol_reader/protocol_reader.py
index 309a25cd8b3..9d4a47f4a5c 100644
--- a/api/src/opentrons/protocol_reader/protocol_reader.py
+++ b/api/src/opentrons/protocol_reader/protocol_reader.py
@@ -1,6 +1,6 @@
 """Read relevant protocol information from a set of files."""
 from pathlib import Path
-from typing import Optional, Sequence
+from typing import Optional, Sequence, Dict, Union
 
 from opentrons.protocols.parse import PythonParseMode
 
@@ -24,6 +24,7 @@
     JsonProtocolConfig,
     PythonProtocolConfig,
 )
+from ..protocol_engine.types import RunTimeParameterValues
 
 
 class ProtocolReader:
@@ -53,7 +54,10 @@ def __init__(
         self._file_hasher = file_hasher or FileHasher()
 
     async def save(
-        self, files: Sequence[BufferedFile], directory: Path, content_hash: str
+        self,
+        files: Sequence[BufferedFile],
+        directory: Path, content_hash: str,
+        run_time_param_values: RunTimeParameterValues,
     ) -> ProtocolSource:
         """Compute a `ProtocolSource` from buffered files and save them as files.
 
@@ -65,6 +69,7 @@ async def save(
             files: List buffered files. Do not attempt to reuse any objects
                 in this list once they've been passed to the ProtocolReader.
             directory: Name of the directory to create and place files in.
+            run_time_param_values: Client-supplied values for Run Time Parameters.
 
         Returns:
             A ProtocolSource describing the validated protocol.
@@ -98,6 +103,7 @@ async def save(
             config=self._map_config(role_analysis),
             robot_type=role_analysis.main_file.robot_type,
             metadata=role_analysis.main_file.metadata,
+            run_time_param_values=run_time_param_values,
         )
 
     async def read_saved(
@@ -164,6 +170,9 @@ async def read_saved(
             config=self._map_config(role_analysis),
             robot_type=role_analysis.main_file.robot_type,
             metadata=role_analysis.main_file.metadata,
+            # We are not passing any RTP values, just reading the existing protocol.
+            # RTPs passed when the protocol was uploaded/ analyzed will be in the analysis.
+            run_time_param_values=None,
         )
 
     @staticmethod
diff --git a/api/src/opentrons/protocol_reader/protocol_source.py b/api/src/opentrons/protocol_reader/protocol_source.py
index ab1aa21e375..7fbfc60a241 100644
--- a/api/src/opentrons/protocol_reader/protocol_source.py
+++ b/api/src/opentrons/protocol_reader/protocol_source.py
@@ -5,6 +5,7 @@
 from typing import Any, Dict, List, Optional, Union
 from typing_extensions import Literal
 
+from opentrons.protocol_engine.types import RunTimeParameterValues
 from opentrons.protocols.api_support.types import APIVersion
 
 from opentrons_shared_data.robot.dev_types import RobotType
@@ -122,3 +123,4 @@ class ProtocolSource:
     metadata: Metadata
     robot_type: RobotType
     config: ProtocolConfig
+    run_time_param_values: Optional[RunTimeParameterValues]
diff --git a/api/src/opentrons/protocol_runner/legacy_wrappers.py b/api/src/opentrons/protocol_runner/legacy_wrappers.py
index 6a816f5e9a1..a469809e411 100644
--- a/api/src/opentrons/protocol_runner/legacy_wrappers.py
+++ b/api/src/opentrons/protocol_runner/legacy_wrappers.py
@@ -94,6 +94,7 @@ def read(
             extra_data={
                 data_path.name: data_path.read_bytes() for data_path in data_file_paths
             },
+            run_time_param_values=protocol_source.run_time_param_values,
             python_parse_mode=python_parse_mode,
         )
 
diff --git a/api/src/opentrons/protocols/parse.py b/api/src/opentrons/protocols/parse.py
index 712b4fe4416..6d92d31032c 100644
--- a/api/src/opentrons/protocols/parse.py
+++ b/api/src/opentrons/protocols/parse.py
@@ -212,6 +212,7 @@ def _parse_json(
 def _parse_python(
     protocol_contents: Union[str, bytes],
     python_parse_mode: PythonParseMode,
+    run_time_param_values: Optional[Dict[str, Union[float, bool, str]]],
     filename: Optional[str] = None,
     bundled_labware: Optional[Dict[str, "LabwareDefinition"]] = None,
     bundled_data: Optional[Dict[str, bytes]] = None,
@@ -273,6 +274,7 @@ def _parse_python(
         bundled_data=bundled_data,
         bundled_python=bundled_python,
         extra_labware=extra_labware,
+        run_time_param_values=run_time_param_values,
     )
 
     return result
@@ -303,6 +305,7 @@ def _parse_bundle(
 
 def parse(
     protocol_file: Union[str, bytes],
+    run_time_param_values: Optional[Dict[str, Union[float, bool, str]]],
     filename: Optional[str] = None,
     extra_labware: Optional[Dict[str, "LabwareDefinition"]] = None,
     extra_data: Optional[Dict[str, bytes]] = None,
@@ -314,6 +317,8 @@ def parse(
 
     :param protocol_file: The protocol file, or for single-file protocols, a
                         string of the protocol contents.
+    :param run_time_param_values: Values of run-time parameters sent by the client,
+                     to be used during analysis/ run.
     :param filename: The name of the protocol. Optional, but helps with
                      deducing the kind of protocol (e.g. if it ends with
                      '.json' we can treat it like json)
diff --git a/api/src/opentrons/protocols/types.py b/api/src/opentrons/protocols/types.py
index 273a3e877d4..075e1e3ac49 100644
--- a/api/src/opentrons/protocols/types.py
+++ b/api/src/opentrons/protocols/types.py
@@ -74,6 +74,8 @@ class PythonProtocol(_ProtocolCommon):
     bundled_python: Optional[Dict[str, str]]
     # this should only be included when the protocol is not a zip
     extra_labware: Optional[Dict[str, "LabwareDefinition"]]
+    # TODO: Use a common type definition between engine's RTP type & this one
+    run_time_param_values: Optional[Dict[str, Union[float, bool, str]]]
 
 
 Protocol = Union[JsonProtocol, PythonProtocol]
diff --git a/robot-server/robot_server/protocols/router.py b/robot-server/robot_server/protocols/router.py
index e71be06864f..8c2bf36cd45 100644
--- a/robot-server/robot_server/protocols/router.py
+++ b/robot-server/robot_server/protocols/router.py
@@ -1,4 +1,5 @@
 """Router for /protocols endpoints."""
+import json
 import logging
 from textwrap import dedent
 from datetime import datetime
@@ -165,6 +166,10 @@ async def create_protocol(
             " protocol resources on the robot."
         ),
     ),
+    runTimeParameterValues: Optional[str] = Form(
+        default=None,
+        description="Key value pairs of run-time parameters defined in a protocol."
+    ),
     protocol_directory: Path = Depends(get_protocol_directory),
     protocol_store: ProtocolStore = Depends(get_protocol_store),
     analysis_store: AnalysisStore = Depends(get_analysis_store),
@@ -184,6 +189,7 @@ async def create_protocol(
     Arguments:
         files: List of uploaded files, from form-data.
         key: Optional key for client-side tracking
+        runTimeParameterValues: Key value pairs of run-time parameters defined in a protocol.
         protocol_directory: Location to store uploaded files.
         protocol_store: In-memory database of protocol resources.
         analysis_store: In-memory database of protocol analyses.
@@ -205,10 +211,12 @@ async def create_protocol(
         assert file.filename is not None
     buffered_files = await file_reader_writer.read(files=files)  # type: ignore[arg-type]
 
+    parsed_rtp = json.loads(runTimeParameterValues) if runTimeParameterValues else None
     content_hash = await file_hasher.hash(buffered_files)
     cached_protocol_id = protocol_store.get_id_by_hash(content_hash)
 
-    if cached_protocol_id is not None:
+    if not parsed_rtp and cached_protocol_id is not None:
+        # Neither a new protocol nor any run-time parameters passed with an existing protocol.
         resource = protocol_store.get(protocol_id=cached_protocol_id)
         analyses = analysis_store.get_summaries_by_protocol(
             protocol_id=cached_protocol_id
@@ -228,7 +236,8 @@ async def create_protocol(
         )
 
         log.info(
-            f'Protocol with id "{cached_protocol_id}" with same contents already exists. returning existing protocol data in response payload'
+            f'Protocol with id "{cached_protocol_id}" with same contents already exists.'
+            f' Returning existing protocol data in response payload.'
         )
 
         return await PydanticResponse.create(
@@ -243,6 +252,7 @@ async def create_protocol(
             files=buffered_files,
             directory=protocol_directory / protocol_id,
             content_hash=content_hash,
+            run_time_param_values=parsed_rtp,
         )
     except ProtocolFilesInvalidError as e:
         raise ProtocolFilesInvalid(detail=str(e)).as_error(