2000
DOI: 10.1128/aem.66.11.4758-4763.2000
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Identification of a Universally Primed-PCR-Derived Sequence-Characterized Amplified Region Marker for an Antagonistic Strain of Clonostachys rosea and Development of a Strain-Specific PCR Detection Assay

Abstract: We developed a PCR detection method that selectively recognizes a single biological control agent and demonstrated that universally primed PCR (UP-PCR) can identify strain-specific markers. Antagonistic strains of Clonostachys rosea (syn. Gliocladium roseum) were screened by UP-PCR, and a strain-specific marker was identified for strain GR5. No significant sequence homology was found between this marker and any other sequences in the databases. Southern blot analysis of the PCR product revealed that the marker… Show more

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Cited by 68 publications
(66 citation statements)
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“…Random amplified polymorphic DNA (RAPD) has been used in many studies. It is based on the use of short general primers that anneal to unspecified regions in the template DNA whereas universally primed (UP) PCR is based on longer general primers and a higher annealing temperature which makes it more robust in terms of reproducibility (Bulat et al, 1998;Bulat et al, 2000;Lubeck et al, 1999;Sabu et al, 2011). UP-PCR has been used to separate sympatric isolates of Beauveria in Denmark and was used to place isolates in genetic groups (Meyling and Eilenberg, 2006).…”
Section: Elucidation On Molecular Studies Of Entomopathogenic Fungimentioning
confidence: 99%
“…Random amplified polymorphic DNA (RAPD) has been used in many studies. It is based on the use of short general primers that anneal to unspecified regions in the template DNA whereas universally primed (UP) PCR is based on longer general primers and a higher annealing temperature which makes it more robust in terms of reproducibility (Bulat et al, 1998;Bulat et al, 2000;Lubeck et al, 1999;Sabu et al, 2011). UP-PCR has been used to separate sympatric isolates of Beauveria in Denmark and was used to place isolates in genetic groups (Meyling and Eilenberg, 2006).…”
Section: Elucidation On Molecular Studies Of Entomopathogenic Fungimentioning
confidence: 99%
“…The genome structure of the three isolates and their closest phylogenetic neighbours was examined using universally primed PCRs with primers AS4/AA2M2 (Bulat et al, 2000). The set-up and protocol were as described previously (Carlsohn et al, 2007).…”
Section: The Genus Kribbellamentioning
confidence: 99%
“…Universally primed PCR (UP-PCR) is a DNA fingerprinting technique closely related to the random amplified polymorphic DNA (RAPD) method (Bulat et al 1998). UP-PCR has the ability to generate numerous bands from primarily intergenic, less conserved regions of the genome (Bulat et al 1998).…”
Section: Introductionmentioning
confidence: 99%
“…UP-PCR has the ability to generate numerous bands from primarily intergenic, less conserved regions of the genome (Bulat et al 1998). UP primers are longer (15-21 nt) than RAPD primers (typically 10 nt) and therefore can anneal under more stringent conditions (52-60 C) ensuring greater reproducibility of banding profiles.…”
Section: Introductionmentioning
confidence: 99%