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Figure 5 | Plant Methods

Figure 5

From: De novo assembly of potential linear artificial chromosome constructs capped with expansive telomeric repeats

Figure 5

DNA composition of the AC constructs. (A) PFGE of AC constructs. (B) Southern blot hybridized with a telomeric DNA probe. (C) Southern blot hybridized with the rice centromeric satellite repeat CentO. Lane 1: Linear ACs capped with ~2 to 5-kb telomeric DNA at both ends. Lane 2: Mixture of pLL-EHC and back-to-back telomeric DNA without recombination. Note: the large bands associated with pLL-EHC did not hybridize to the telomeric DNA probe in (B). Lane 3: Linear molecules derived from recombination between pLL-EHC and linearized pLL-BKE (~10-kb of non-telomeric DNA, see Experimental Procedures). The arrowhead in (A) points to the non-recombined pLL-BKE molecules. Lane 4: pLL-EHC DNA after incubation with BP clonase. Lane 5: Linear AC constructs capped with ~5 to 10-kb telomeric DNA at the left arm only after recombination between pLL-EHC and expansive telomeric DNA from pLL-TBS. The red square in (B) includes the non-recombined telomeric DNA molecules. Lane 6: Linear AC constructs capped with ~5 to 10-kb telomeric DNA at the right arm only after recombination between pLL-EHC and expansive telomeric DNA from pLL-TSB. The red square in (B) includes the non-recombined telomeric DNA molecules.

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