AScall

AScall performs automated processing and analysis of allele-specific real-time PCR (AS-qPCR) data to call sample genotypes and detect allele-specific genetic variation.

Key Features:

• Automated Data Processing: Automatic processing of large-scale AS-qPCR datasets with support for multiplexing up to 480 reactions per 96-well plate.
• Universal Data Import: Batch import of real-time PCR data in RDML format.
• Amplification Curves Preprocessing: Initial preprocessing of amplification curves to prepare data for downstream analysis.
• Quality Control: Automated checks to validate data integrity and reliability.
• Sample Genotype Calling: Determination of genotypes based on allele-specific amplification signals.
• Results Visualization: Generation of detailed visualizations for interpretation of amplification and genotype data.
• Report Generation: Production of detailed analytical reports summarizing experiment results.
• High-throughput Performance: Demonstrated capability to process large datasets (example: 4,800 qPCR reactions processed in 1.5 minutes as reported).

Scientific Applications:

• Minor Histocompatibility Antigen (MiHA) Genotyping: Detection of donor–recipient mismatches in MiHA studies following allogeneic hematopoietic stem cell transplantation.
• Genetic Variation Detection: Identification of single nucleotide polymorphisms (SNPs), insertions, and deletions using allele-specific primers and real-time PCR.
• High-throughput Allele-specific Genotyping: Large-scale AS-qPCR experiments for research requiring rapid allele discrimination across many samples.

Methodology:

AScall imports RDML files, preprocesses amplification curves, performs automated quality control, and applies an algorithm that interprets fluorescent signals from allele-specific real-time PCR to call sample genotypes.