Agilent 2100 Bioanalyzer Nested Multiplex PCR for GMO Detection Application Note

About this manual

The document details a method for detecting DNA from genetically modified organisms (GMOs) in food samples. It utilizes a nested multiplex PCR assay based on the Biosmart Allin 1.0 GMO Screening System from Promega, which targets the 35S promoter, soya lectin, corn zein, and an internal control.

The procedure evaluates different DNA extraction protocols (CTAB, DNeasy, Wizard Genomic) to determine optimal yield and purity before PCR analysis. The resulting PCR products are analyzed using the Agilent 2100 bioanalyzer with a DNA 500 LabChip or via gel electrophoresis for comparison.

What's inside

• Introduction
• DNA Extraction Protocols
• CTAB Extraction Protocol
• PCR Protocol
• Multiplex PCR Protocol
• Analysis of PCR Products
• Results: DNA Extraction
• Results: Analysis of PCR Products

Frequently asked questions

What genetic elements does the multiplex PCR assay detect?

The assay detects sequences for the 35S promoter, soya lectin (Cectin), corn zein, and an internal positive control.

Which DNA extraction protocol was found to have the highest yield and purity?

The CTAB extraction protocol showed the highest levels of DNA recovery with a high DNA/protein ratio.

What are the base pair sizes of the four PCR products generated?

The products correspond to 118 bp (Soy lectin), 153 bp (35S Promoter), 217 bp (Internal control), and 278 bp (Corn zein).