GENEQUALITY^® CFTR

Description

GENEQUALITY^® CFTR is an in vitro diagnostic kit for amplification-based library preparation, designed for comprehensive analysis of the CFTR gene using Next-Generation Sequencing (NGS).
The kit enables reliable detection of mutations, indels, and CNVs in exonic regions, exon–intron boundaries, promoter regions, and clinically relevant intronic variants (including poly-T and TG polymorphisms).

Product Characteristics

• Low Input DNA: only 10 ng per reaction.
• Illumina Compatibility: paired-end sequencing 2×150 bp.
• Fast Workflow: complete protocol in under 3 hours, with less than 45 minutes of hands-on time.
• One-Tube Library Prep: reduces contamination risk and simplifies sample handling.
• Comprehensive CFTR Coverage: exons, introns, regulatory regions, poly-T and TG repeats.
• CNV Analysis: direct identification of mutations across all coding regions.
• Validated Sample Types: whole blood (EDTA), amniotic fluid, buccal swab, and Dried Blood Spots (DBS).

Kit content

• The device contains all the reagents required for genomic library preparation.

Further Information

GENEQUALITY^® CFTR enables library preparation from high-quality genomic DNA isolated from whole blood (EDTA), amniotic fluid, and Dried Blood Spots (DBS).
The kit provides reagents for the preparation and purification of amplicon libraries for CFTR gene analysis.
GENEQUALITY^® CFTR uses high-multiplex PCR for target region amplification.
The resulting libraries are compatible with sequencing on Illumina platforms.

The CFTR amplicon library preparation procedure consists of the following steps:

1. Target region amplification of the CFTR gene: 10 ng of purified genomic DNA (gDNA) is amplified using high-multiplex PCR. The entire reaction is performed in a single tube per sample. Each primer includes, at the 5’ end, a recognition sequence serving as a binding site for indexed adapters introduced in the second PCR.
2. Amplificate dilution: The product of the first amplification is diluted before being used as a template in the second PCR. This dilution removes potential inhibitors (e.g., salts, primers, residual enzymes) and adjusts DNA concentration to an optimal level, improving specificity and efficiency of the subsequent reaction.
3. Amplification with UDI adapters: The diluted products of the first amplification undergo a second PCR using library adapters containing unique dual indexes (UDIs). These adapters allow unique identification of each sample via the indices and provide the sequences required for cluster formation during sequencing.
4. Pooling and Purification: The indexed libraries are pooled into a single mix. The pool is then purified using magnetic beads and yield is assessed by fluorometric quantification.
5. Sequencing: The pooled libraries are ready for sequencing on a compatible Illumina instrument.

Ordering Information