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Chimerism in PCR products from a multigene family
D Bhavsar1, H Zheng, J Drysdale
1Department of Biochemistry, Tufts Medical School, Boston, MA 02111.
Biochemical and Biophysical Research Communications
|November 30, 1994
Summary
Polymerase chain reaction (PCR) amplification of multigene families can create artificial chimeric sequences. This study identifies a potential hazard where PCR may combine different gene sequences into one, affecting genetic analysis.
Area of Science:
- Molecular Biology
- Genetics
- Bioinformatics
Background:
- Multigene families are common in genomes.
- Polymerase chain reaction (PCR) is widely used for DNA amplification.
- Conserved sequences are often targeted for PCR primers.
Purpose of the Study:
- To describe a potential hazard in sequencing multigene families using PCR.
- To identify the formation of chimeric sequences during PCR.
Main Methods:
- Amplification of mixed DNA populations from a multigene family.
- Use of conserved primers for PCR.
- Analysis of PCR products.
Main Results:
- A potential hazard was identified in PCR-based sequencing of multigene families.
- A chimeric product was observed, seemingly formed from two related sequences during PCR.
- This artifact can complicate the accurate sequencing of related genes.
Conclusions:
- PCR amplification of mixed populations from multigene families carries a risk of generating chimeric sequences.
- Researchers must be aware of this potential artifact when analyzing gene families.
- Careful analysis is needed to distinguish true sequences from PCR-induced chimeras.