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We analysed this parasite population by Southern blot, using a probe diagnostic for the specific integration event.
Cells were selected with G418, and 96 resistant clones were screened by southern blot using a probe specific for the Flag-SMAD1 sequence.
Detection of siRNAs was carried out by a northern blot using a probe specific for pericentromeric dg repeats.
We performed genomic Southern blot using a probe near the insertion site of P26 (Fig. 5 C).
Moreover, a Southern blot using a probe for MAT1-2-1 identified the remaining four isolates as being identical to CCF3942.
To test this, genomic DNA from Clone A1, and Clone A12 included for comparison, was analysed by Southern blot using a probe for the GFP gene.
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Currently, the number of D4Z4 repeats is usually determined by Southern blotting using a probe that hybridizes to the centromeric flanking sequence, p13E-111.
Northern blotting using a probe for the 3' flanking region of the GSA target showed no signal in either the 35S GS-IR or the CoYMV:GS-IR transgenic line (Figure S4), although both exhibited unequivocal PTGS (Figure S2a, S2b).
We next carried out Southern blotting using a probe that targets the 30 portion of the LUC coding sequence.
Two clones were selected following further screening by Southern blotting using a probe located 5' to the targeting vector as described previously [ 26].
DNA was then isolated, digested with SacI, and the fragments analyzed by southern blotting using a probe corresponding to bp +1060 to +1264 relative to the transcription start site.
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