Propagating (Connecting) Runs across Steps

 Creating run connections is an important feature of Riffyn. When runs are connected across steps, the data across these steps will be automatically joined together on data export, allowing a comprehensive cause/effect analysis to be performed across an entire process. 

A run can be created on a step by propagation of existing runs of an upstream step to a downstream stepWhen runs are propagated from one step to another they are automatically connected across these steps. If runs are created manually or via file upload they will not be automatically connected across steps. 

In the HPLC of Cell Culture Supernatant public process, connecting runs enables assessment of the impact of the centrifugation temperature used to generate cell culture supernatant (data generated on "Centrifuge Samples" step) on the concentration of acetate found in this supernatant (data generated on "Inject Samples" step). If runs are created for these two steps but not connected, then the data across these steps will not be joined, and this cause/effect analysis will not be possible.  

To demonstrate how to propagate runs, use the Public Process HPLC of Cell Culture Supernatant. In this demo, step "Prep Mobile Phases" has one run (named "Mobile Phase A"). All of the following examples show the propagation of a run from an upstream step to downstream step(s), but it is important to note that propagation from a downstream step to an upstream step is also possible.

 Example 1 - Propagating a run from one step to the next with a 1:1 split

A connection from a run in one step to a run in another can be achieved with a "1:1 split", meaning each run on the first step is connected to one and only one run on the next step.

For example, a batch of runs could be created for the "Prep Mobile Phase" step, each of which should have a single run in the "Column Equilibration" step. In that case, runs should be propagated from the "Prep Mobile Phases" step to the "Column Equilibration" step:

  1. In the Flow Diagram, click on a step with run(s) on it; in this example, "Prep Mobile Phases" has one run on it called "Mobile Phase A". In the Bottom Panel, navigate to the "1. Plan Experiment" tab. Notice that a yellow anchor symbol appears in the lower left corner of the "Prep Mobile Phases" flow diagram box. 

  2. Click the anchor on the step to which you'd like to propagate the runs (in this example, "Column Equilibration").



  3. The connected steps turn blue in the Flow Diagram, and the Bottom Panel switches to "Flow Mode" (showing two steps side by side).

  4. Select the runs on "Prep Mobile Phases" to be propagated to "Column Equilibration". Note that individual runs can be chosen by clicking the box next to the run, or all runs can be selected by checking the box to the left of the "Label" Header.



  5. Navigate to the Toolbar on the bottom of the screen.
  6. Choose the split ratio. The split ratio indicates how many runs to create in the connected step. The default is 1:1 (meaning one run will be created in the connected step for every one run in the original step). In this example we will use a split ratio of 1:1. For different split ratios, see Example 2.
  7. Click the "Go" button to create the connected run.

  8. "Column Equilibration" will now have a run on it.

  9. To view the run connection(s), select a run by checking the box next to a run. In this example, the run on "Prep Mobile Phases" is selected. Notice that the run on step "Column Equilibration" is highlighted in blue, indicating that these runs are connected.



  10. Notice that the Mobile Phase output in "Column Equilibration" (GR-DvAx), has been assigned as the Mobile Phase input of the run on "Column Equilibration".  





 

 

Example 2 - Propagating a run from one step to the next with a 1:N split

A connection from a run in one step to many runs in another can be achieved with a "1:N split", meaning each run on the first step is connected to more than one runs on the next step. 

For example, a batch of runs could be created for the "Prep Mobile Phase" step, each of which should have a multiple runs in the "Column Equilibration" step. A 1:3 split of runs should be propagated from the "Prep Mobile Phases" step to the "Column Equilibration" step as follows:

  1. Follow tasks 1-7 as outlined in Example 1, except in task #6, instead of selecting a 1:1 ratio, select a 1:3 ratio. Click "Go".



  2. See that "Column Equilibration" now has 3 runs on it, each of which is connected to the run on "Prep Mobile Phases" (as evidenced by the blue highlighting that appears on the "Column Equilibration" runs when the run on "Prep Mobile Phases" selected).


     

Example 3 - Propagating a run from one step to multiple steps with a 1:1 split

A connection from a run in one step to a run in multiple other steps can be achieved with a "1:1 split", meaning each run on the first step is connected to one and only one run on multiple steps.

For example, a batch of runs could be created for the "Prep Mobile Phase" step, each of which should have a single run in the "Column Equilibration" step. In that case, runs should be propagated from the "Prep Mobile Phases" step to the "Column Equilibration" step:

For example, a batch of runs could be created for the "Prep Mobile Phase" step, each of which should have a single run in the "Column Equilibration" step. A sample is then injected in the "Inject Sample" step after "Column Equilibration". Therefore a run on the "Prep Mobile Phases" step needs to be connected to a run on the "Column Equilibration" step, and subsequently each run in the "Column Equilibration" step should be be connected to a run on the "Inject Samples" step. Rather than propagating these runs by performing 2 propagations, you can create connected runs on both the "Column Equilibration" and "Inject Sample" steps at the same time, by propagating runs from "Prep Mobile Phases" to "Inject Sample" through "Column Equilibration":

  1. Click on "Prep Mobile Phases", navigate to the "1. Plan Experiment" tab, and notice that a yellow anchor symbol appears in the lower left corner of the "Prep Mobile Phases" flow diagram box. (See Example 1, Task 1).

  2. Click the anchor on the furthest upstream or downstream step to which you'd like to propagate runs (in this example, "Inject Sample").

  3. Notice that all connected steps along the default "Run Path" turn blue in the Flow Diagram, and the Bottom Panel switches to "Flow Mode" (showing "Prep Mobile Phases" and "Inject Sample" side-by-side).



  4. Select the runs on "Prep Mobile Phases" for propagation to "Inject Samples" through "Column Equilibration". Note that individual runs can be chosen by clicking the box next to the run, or all runs can be selected by checking the box to the left of the "Label" Header.

  5. See/Select the "Run Path" on the bottom toolbar "Create Connected Runs from "Prep Mobile Phases" to  "Inject Samples" via..." The default selection is "Column Equilibration", which creates the run path "Prep Mobile Phases" -> "Column Equilibration" -> "Inject Samples". As such, all of these steps are highlighted in blue in the Process Flow diagram. Since the objective is to propagate and connect runs to both "Column Equilibration" and "Inject Samples", this is the desired run path. 



  6. Next, choose the split ratio. In this example a split ratio of 1:1 is chosen. For different split ratios, see Example 4.

  7. Select the step after which runs will be split. In this example, the options are "Prep Mobile Phases" and "Column Equilibration", however, since it's a 1:1 split, this command is irrelevant. For an example of how to when to change the "Split after" command, see Example 4.

  8. Click the "Go" button to create and connect the runs.

  9. Notice that both steps "Inject Samples" and "Column Equilibration" now have runs on them. The run on "Prep Mobile Phases" is connected to the run on step "Column Equilibration", which in turn is connected to the run on step "Inject Samples". That is, the run on step "Inject Samples" is connected the run on step "Prep Mobile Phases" via the run on step "Column Equilibration". To view run connections, select a step in the Process Flow Diagram, then click the anchor on another step in the Process Flow diagram. This will show the Run Flow on the Bottom Panel. Select run(s) on one of the steps; run(s) highlighted in blue on the other step are connected to this run.  

    Run on "Inject Samples" is created and connected to step "Prep Mobile Phases" via a run on step "Column Equilibration".


    Run on step "Column Equilibration" is created and connected to step "Prep Mobile Phases".

 

Example 4 - Propagating a run from one step to multiple steps with a 1:N split

 

A connection from a run in one step to many runs in multiple steps can be achieved with a "1:N split", meaning each run on the first step is connected to more than one runs on the other steps. 

For example, a batch of runs could be created for the "Prep Mobile Phase" step, each of which should have a single run in the "Column Equilibration" step. 3 samples are then injected in the "Inject Sample" step after "Column Equilibration". Therefore a run on the "Prep Mobile Phases" step needs to be connected to a run on the "Column Equilibration" step, and subsequently each run in the "Column Equilibration" step should be be connected to 3 runs on the "Inject Samples" step:

  1. Follow tasks 1-5 in Example 3. 

  2. Next, choose the split ratio. Because we want one run in step "Column Equilibration" to be connected to 3 runs on step "Inject Samples", choose a split ratio of 1:3.

  3. Select the step after which runs will be split. Because a 1:1 ratio is needed from step "Prep Mobile Phases" to step "Column Equilibration" (one batch of Mobile Phase is connected with one "Column Equilibration" run), but a 1:3 ratio is needed from step "Column Equilibration" to step "Inject Samples" (one "Column Equilibration" run is connected to three Inject Samples runs), it is necessary to split the runs after step "Column Equilibration". 

  4. Click the "Go" button to create and connect the runs.

  5. Notice that 3 runs are created on step "Inject Samples", which are connected to step "Prep Mobile Phases", as shown by the blue highlighting that appears on these runs when the run on step "Prep Mobile Phases" is selected. The runs on step "Inject Samples" are connected to step "Prep Mobile Phases" via step "Column Equilibration" as shown by blue highlighting on the Process Flow Diagram.



    However, only one run is created on step "Column Equilibration". This one run on step "Column Equilibration" is connected to one run on step "Prep Mobile Phases", but to three runs on step "Inject Samples". (To see run connections, select a step in the Process Flow Diagram, then click the anchor on another step in the Process Flow diagram. This will show you the Run Flow on the Bottom Panel. Select run(s) on one of the steps; run(s) highlighted in blue on the other step are connected to this run.)

    Only one run is created on step "Column Equilibration", which is connected to one run on step "Prep Mobile Phases".


    This one run on step "Column Equilibration" is connected to three runs on step "Inject Samples".
     

 

Example 5 - Changing the Run Path when propagating runs

When an output of a step is connected to more than one downstream step, a choice exists as to which run path to use when propagating runs. For example, in the Public Process "HPLC of Cell Culture Supernatants", step "Prep Mobile Phases" has a single output, "Mobile Phase", which is connected to both step "Column Equilibration" and step "Inject Samples". When you propagate runs from "Prep Mobile Phases" to "Inject Samples", you can either propagate runs through "Column Equilibration" (meaning, runs on "Inject Samples" will be connected to runs on "Prep Mobile Phases" via runs on "Column Equilibration") or you can propagate runs directly from "Prep Mobile Phases" to "Inject Samples" and bypass "Column Equilibration". Example 3 demonstrated how to propagate runs from "Prep Mobile Phases" to "Inject Samples" via "Column Equilibration". This example demonstrates how to propagate runs from "Prep Mobile Phases" directly to "Inject Samples", skipping "Column Equilibration". Two use cases exist; if a batch of Mobile Phase skips the "Column Equilibration" step and goes straight to injecting samples; or if one batch of Mobile Phase is used for "Column Equilibration" and another batch is used for sample injection. 

  1. Follow Example 3 instruction tasks 1-4. Notice that step "Prep Mobile Phases", "Column Equilibration", and "Inject Samples" are all highlighted in blue in the Process Flow Diagram, indicating a default run path from "Prep Mobile Phases" to "Inject Samples" is through "Column Equilibration".

  2. Change the run path to connect "Prep Mobile Phases" directly to "Inject Samples" by choosing the "blank" selection from the dropdown menu next to "Create Connected Runs from "Prep Mobile Phases" to  "Inject Samples" via..." 
     
  3. Notice that the run path changes in the Process Flow Diagram. "Column Equilibration" turns gray, while Steps "Prep Mobile Phases" and "Inject Samples" remain blue, indicating that newly propagated runs from "Prep Mobile Phases" will bypass step "Column Equilibration" and flow directly to "Inject Samples".

 Also see How to decipher run connections in previously created experiments.

Was this article helpful?
0 out of 0 found this helpful
Have more questions? Submit a request