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PCR-generated padlock probes detect single nucleotide variation in genomic DNA
D O Antson1, A Isaksson, U Landegren
1Rudbeck Laboratory, Department of Genetics and Pathology, SE-751 85, Uppsala, Sweden and Department of Molecular Cell Biology, Leiden University Medical Center, Wassenaarseweg 72, 2333 AL Leiden, The Netherlands.
Nucleic Acids Research
|June 28, 2000
Summary
Padlock probes enable genetic investigations, but are hard to synthesize. A new PCR-based method allows flexible, enzymatic synthesis of these probes for improved genetic analysis.
Area of Science:
- Molecular Biology
- Genetics
- Biotechnology
Background:
- Padlock probes are circularizing oligonucleotides with potential in genetic analysis.
- Standard chemical synthesis poses challenges for producing long, functional padlock probes.
Purpose of the Study:
- To develop a flexible and efficient method for synthesizing padlock probes.
- To demonstrate the utility of enzymatically synthesized padlock probes for genetic variation analysis.
Main Methods:
- A PCR-based protocol for small-scale enzymatic synthesis of padlock probes.
- Enzymatic synthesis allows for longer probes densely labeled with detectable functions.
Main Results:
- The protocol facilitates the flexible synthesis of padlock probes.
- Probes synthesized via this method show increased detection signals.
- Demonstrated utility in analyzing single nucleotide variations in human DNA.
Conclusions:
- The described PCR-based enzymatic synthesis offers an advantageous alternative to chemical synthesis for padlock probes.
- This method enhances the capability for genetic investigations using padlock probes.