APAgene™ GOLD Genome Walking Kit

Temperature-stable APAgene™ GOLD Genome Walking Kits

Greater convenience and lower shipping costs for everyone.

Our international customers will particularly benefit from the ability to ship without requiring expensive cold chain logistics. Order today and save on shipping—a major benefit for our international customers

Our kit was featured in Molecular Biotechnology (D. Cullen et al., 11 January 2011 pp. 1-13) where it rated the best amongst our competitors and in Food Chemistry (MA Fraiture et al, 15 March 2014 pp 60-69) as an approach in identifying unauthorised GMOs

Ultimate tool for genomic research

APAgene™ GOLD Genome Walking Kits use our patented APA technology to help you rapidly (in just one day!) and reliably amplify unknown genomic DNA. Powered by our patented APA technology, you can amplify and isolate sequences flanking transgenes, STSs, and ESTs in just 24 hours.

• Determine unknown genomic regions: Easily isolate and sequence localized DNA that flanks a known sequence from large clones (like P1, YAC, and BAC DNA) or whole genomic DNAs.

• Sequence clone insert ends: Quickly analyze the terminal sequences of large genomic clones, such as BAC, YAC, and P1 vectors.

• Identify gene regulatory elements: Isolate and characterize the unknown 5' promoter control regions and 3' transcriptional terminators of cDNAs.

• Perform rapid amplification of cDNA ends (RACE): Efficiently isolate the unknown 5' and 3' RACE products from your first-strand cDNA

Amplification process

To successfully amplify the flanking sequences, you'll typically need to use a set of three nested GSPs (GSPa, GSPb, and GSPc) in the reaction. Our patented technology is powered by a combination of two key primer types:

1. Our proprietary Degenerate Random Tagging (DRT) primers.
2. Your own Gene-Specific Primers (GSPs).

Our Degenerate Random Tagging (DRT) primers are universal binding primers engineered with three essential components that drive the amplification process:

• A Tagging Sequence: For specific downstream amplification.

• A Random Sequence: To allow broad, non-specific binding across the genome.

• A Degenerate Sequence: To enhance binding affinity to various DNA templates.

How it Works: The Mechanism Behind the Success

The core of the "walking" reaction is the binding of our DRT primers to the unknown flanking sequence during the primary PCR. All DRT primers have a shared tagging sequence, but their different random sequences enable widespread, non-specific binding across the genome. This powerful binding, combined with our patented DRT primers, optimized cycling conditions, and unique PCR buffer, dramatically enhances the target amplification process. Furthermore, we utilize UAP primers—specialized hairpin-containing primers—which significantly suppress non-specific amplification, leading to a much cleaner and more accurate final product.

Other publications:

- An integrated strategy involving high-throughput sequencing to characterize an unknown GM wheat event in Canada

- DNA walking strategy to identify unauthorized genetically modified bacteria in microbial fermentation products

- ALF: a strategy for identification of unauthorized GMOs in complex mixtures by a GW-NGS method and dedicated bioinformatics analysis

- Development and validation of an integrated DNA walking strategy to detect GMO expressing cry genes

- Molecular characterization for food safety assessment of a genetically modified late blight resistant potato

- Comparison of DNA Walking Methods for Isolation of Transgene-Flanking Regions in GM Potato

- Palindromic sequence-targeted (PST) PCR: a rapid and efficient method for high-throughput gene characterization and genome walking