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. 2012 Sep 12;40(21):10866–10877. doi: 10.1093/nar/gks859

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© The Author(s) 2012. Published by Oxford University Press.

This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/3.0/), which permits unrestricted, distribution, and reproduction in any medium, provided the original work is properly cited.

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Figure 4. — Class B products form with high frequency. (A) A cPCR assay to detect class B products. The indicated primers (in red) were used for colony PCR reactions to specifically amplify products in strains containing class B-I or class B-II circles. (B) The cPCR assay is sensitive and specific for class B products. Original class B strains and strains converted to His⁺ by class B genomic DNA transformation (Figure 2C and D) were subjected to the cPCR assay. (C) His⁺ phenotype tracks with the class B circle. A His⁺ stability experiment (described in Figure 2E) was performed on parent strains transformed with a class B product. Colonies that lost the His⁺ phenotype (YPD, His⁻) and rare colonies that retained the phenotype (YPD, His⁺) were tested by the cPCR assay for the presence of a class B episome. (D) Class B products constitute the majority of Zorro3 retrotransposition events. For two strains (containing Zorro3mHIS3AI or Zorro3pA⁻mHIS3AI), 44 independently derived retrotransposition events (all unlabelled lanes) were tested by cPCR for the presence of class B-I or class B-II products. 13 = an original class B-I clone (control). 14 = an original class B-II clone (control).