Sickle Cell Monochrome Multiplex qPCR Data Analysis Suite

Application Description

This analytical suite provides a standardized framework for the high-throughput interpretation of monochrome multiplex qPCR assay, tailored for sickle cell anemia genotyping. By integrating automated Cycle Threshold (Cq) delta-analysis, the suite enables high-precision discrimination between HbA (Normal) and HbS (Sickle) allele. The platform incorporates a dual-signal of monochrome multiplexing qPCR protocol to ensure clinical-grade reliability by cross-referencing internal controls for DNA quality and PCR inhibition.

Pre-analysis: PCR cycle (X)

Enter the PCR cycle index X used in your formulation. This value is required (integer) before you upload Signal 1 and is carried through later calculation steps.

Suite Data Entry: Signal 1

Save X (PCR cycle) above to enable file upload and analysis.

Use the standard export with columns Well, Fluor, Target, Content, Sample, Cq. The suite only reads Sample, Target, and Cq (other columns are ignored). Targets must start with capital N or M (Ct(N) / Ct(M)), at most one of each per sample.

Drag and drop your file here, or browse Spreadsheet formats · .csv, .xlsx, .xls

📌 Suite Input Requirements

Expected header row (typical qPCR export): Well · Fluor · Target · Content · Sample · Cq

Sample	Target	Cq
Lab_Sample_01	N	22.41
Lab_Sample_01	M	31.00

Files must be .xlsx or .csv. Columns Well, Fluor, and Content are kept in the file but not used by the suite.
Signal 1 & Signal 2: Target must start with capital N or M (Ct(N) vs Ct(M)); each sample has at most one N-row and one M-row.
Non-detects (N/A) are automatically normalized to 31.0.

Suite Data Entry: Signal 2

Upload and validate Signal 1 first.

Upload the quantification export for Signal 2 (second channel).

Drag and drop your file here, or browse Spreadsheet formats · .csv, .xlsx, .xls

📌 Suite Input Requirements

Expected header row (typical qPCR export): Well · Fluor · Target · Content · Sample · Cq

Sample	Target	Cq
Lab_Sample_01	N	22.41
Lab_Sample_01	M	31.00

Files must be .xlsx or .csv. Columns Well, Fluor, and Content are kept in the file but not used by the suite.
Signal 1 & Signal 2: Target must start with capital N or M (Ct(N) vs Ct(M)); each sample has at most one N-row and one M-row.
Non-detects (N/A) are automatically normalized to 31.0.

Session & workflow

Reset options are available on every step. Clear uploads removes files and results but keeps your PCR cycle X. Restart from the beginning clears the full session (including X) and all cached analysis data for this browser session.