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High output genetic mapping of polyploids using PCR-generated markers.

B W Sobral1, R J Honeycutt

  • 1California Institute of Biological Research, 11099 North Torrey Pines Road, Suite 300, 92037, La Jolla, CA, USA.

TAG. Theoretical and Applied Genetics. Theoretische Und Angewandte Genetik
|November 7, 2013
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Summary
This summary is machine-generated.

Optimized polymerase chain reaction (PCR) protocols using the Stoffel fragment and improved thermal profiles enhance genetic mapping and genomic fingerprinting efficiency. This method increases useful primers and polymorphisms while reducing enzyme costs.

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Area of Science:

  • Molecular Biology
  • Genetics
  • Plant Science

Background:

  • Polymerase chain reaction (PCR) with arbitrary primers is effective for genetic mapping and genomic fingerprinting.
  • Existing PCR protocols can be improved for increased productivity in large-scale mapping efforts.

Purpose of the Study:

  • To enhance the efficiency of genetic mapping and genomic fingerprinting.
  • To optimize PCR protocols for increased locus screening and polymorphism detection.

Main Methods:

  • Utilized the Stoffel fragment, a modified Taq polymerase, in PCR reactions.
  • Optimized the thermal cycling profile for PCR.
  • Applied the modified protocol to map the genome of Saccharum spontaneum 'SES 208'.

Main Results:

  • Increased the number of loci screened per primer by 30%.
  • Achieved an 80% increase in polymorphisms detected per primer.
  • Enhanced the usability of primers from 70% to 97% for reproducible fingerprints.
  • Reduced enzyme cost per reaction approximately 1.6-fold.
  • Successfully placed some generated markers into linkage groups.

Conclusions:

  • The modified PCR protocol significantly improves the efficiency and cost-effectiveness of genetic mapping and genomic fingerprinting.
  • The use of the Stoffel fragment and optimized thermal profiles are key to enhanced PCR performance.
  • This optimized method facilitates more robust marker development for polyploid species like sugarcane.