PCR-Free Cloning of Repetitive DNA Sequences
Author Information
Author(s): Annika Scior, Steffen Preissler, Miriam Koch, Elke Deuerling
Primary Institution: University of Konstanz
Hypothesis
Can a PCR-free method be developed for the efficient generation of highly repetitive DNA sequences?
Conclusion
The study presents a PCR-free cloning strategy that allows for the efficient generation of highly repetitive DNA sequences of defined lengths.
Supporting Evidence
- The method allows for the cloning of defined stretches of Poly-Q encoding sequences.
- It can also generate polyadenine sequences for in vitro transcription of mRNA.
- The approach is straightforward and can be completed in a short time frame.
Takeaway
The researchers found a way to clone long repetitive DNA sequences without using PCR, making the process easier and more accurate.
Methodology
The study used antiparallel oligonucleotides flanked by Type IIS endonucleases for seamless elongation of repetitive DNA sequences.
Limitations
The method may not be suitable for all types of repetitive sequences and relies on specific enzyme availability.
Digital Object Identifier (DOI)
Want to read the original?
Access the complete publication on the publisher's website