Required Measurements — qPCR Study

MODEL "Required Measurements — qPCR Study"
DESCRIPTION "A synthetic relative-expression study with required Cq inputs, a worked measurement table, and an explicit missing-data state."
VERSION "1.0.0"
AUTHOR "Grid Team"
TAGS "biology", "research", "qpcr", "validation"

# Enter the synthetic Cq values shown on the sheet, or your own reviewed values.
# Blank defaults keep an unmeasured or cleared input from becoming numeric zero.
input TreatedTargetCq = BLANK
  VALIDATE BETWEEN 1 AND 45 MESSAGE "Enter the measured treated-target Cq within this example's 1–45 cycle window."
input TreatedReferenceCq = BLANK
  VALIDATE BETWEEN 1 AND 45 MESSAGE "Enter the measured treated-reference Cq within 1–45 cycles."
input ControlTargetCq = BLANK
  VALIDATE BETWEEN 1 AND 45 MESSAGE "Enter the measured control-target Cq within 1–45 cycles."
input ControlReferenceCq = BLANK
  VALIDATE BETWEEN 1 AND 45 MESSAGE "Enter the measured control-reference Cq within 1–45 cycles."

# Equal target/reference efficiencies near doubling are an assay assumption.
CqPresent = (
  ISNUMBER(TreatedTargetCq) AND ISNUMBER(TreatedReferenceCq)
  AND ISNUMBER(ControlTargetCq) AND ISNUMBER(ControlReferenceCq)
)
CqValid = (CqPresent
  AND TreatedTargetCq >= 1 AND TreatedTargetCq <= 45
  AND TreatedReferenceCq >= 1 AND TreatedReferenceCq <= 45
  AND ControlTargetCq >= 1 AND ControlTargetCq <= 45
  AND ControlReferenceCq >= 1 AND ControlReferenceCq <= 45
)

output StudyStatus IS string = CqValid THEN "Measurements supplied; review assay assumptions before interpretation." ELSE "Missing or invalid Cq measurement; no relative expression reported."
output DeltaDeltaCq IS number = CqValid THEN QPCR_DELTA_DELTA_CQ(TreatedTargetCq, TreatedReferenceCq, ControlTargetCq, ControlReferenceCq) ELSE #N/A
output RelativeExpression IS number = CqValid THEN 2 ^ (-DeltaDeltaCq) ELSE #N/A
output Assumptions IS string = "Synthetic demonstration; Cq values may summarize technical replicates but do not establish biological replication. The 2^(-delta-delta-Cq) comparison assumes a stable reference and equal efficiencies near doubling. These input windows do not validate an assay."
output ScenarioGuide IS string = "Enter 23, 20, 25, 20 in the corresponding I/O inputs: relative expression is 4. Save the study. Change treated target to 22: expression is 8. Clear any Cq to see the missing state. Reopen the saved archive to restore 4, or reset inputs to restore blank measurements."

A1 = "Synthetic measurement to enter in I/O"
B1 = "Cq (cycles)"
A2 = "Treated target"
B2 = 23
A3 = "Treated reference"
B3 = 20
A4 = "Control target"
B4 = 25
A5 = "Control reference"
B5 = 20
A7 = StudyStatus
A8 = "Relative expression"
B8 = RelativeExpression
A10 = Assumptions
A11 = ScenarioGuide

END MODEL