Suppression of randomly primed polymerase chain reaction products (random amplified polymorphic DNA) in heterozygous diploids.
In a study of genetic polymorphism in the gypsy moth Lymantria dispar we observed the aberrant inheritance of a random amplified polymorphic DNA (RAPD) fragment designated H11-589. This fragment was present in amplification products of F1 progeny of different crosses although it was not amplified from either parental DNA. DNA-mixing experiments revealed that the presence of DNA containing a template for another product (H11-746), amplified with the same primer, suppressed the synthesis of H11-589. The templates for both RAPD products were highly repetitive and scattered throughout the L. dispar genome. Southern hybridization and sequence analysis of H11-746 and H11-589 revealed an extensive sequence homology and an internal repetitive motif of 17 nucleotides present in both products. Interactions between templates for H11-746 and H11-589 are expected to occur during the polymerase chain reaction (PCR), offering an explanation for the suppression of the amplification of H11-589. The role of the internal repetitive motif and of the copy number of both templates in the suppression effect are discussed. Our results corroborate doubts regarding the suitability of the RAPD technique for quantitative genetic analysis, in particular where mixed populations are concerned.
Reineke
A
A
University of Hohenheim, Institute of Phytomedicine, Stuttgart, Germany. areineke@waite.adelaide.edu.au
Karlovsky
P
P
Zebitz
C P
CP
D000818:Animals; D001483:Base Sequence; D004171:Diploidy; D005819:Genetic Markers; D006579:Heterozygote; D008969:Molecular Sequence Data; D009036:Moths / Q000235:genetics*; D016133:Polymerase Chain Reaction; D019105:Random Amplified Polymorphic DNA Technique; D017422:Sequence Analysis, DNA
D005819:Genetic Markers