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225565Supplemental_Figure_S4

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susan-dejesus
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225565Supplemental_Figure_S4

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  1. 1 2 3 4 5 6 7 8 9 10 Supplemental Figure S4: Expression of the ZmISA1 transgene in the Arabidopsisisa1-1 isa2-2mutant host line. Leaf cell RNAswere extracted from the isa1-1 isa2-2host strain expressing ZmIsa1 using the plant RNeasy kit from Qiagen(Courtaboeuf, France). The RT-PCR amplification was performed using the OneStepRT-PCR kit from Qiagen(Courtaboeuf, France). Different amounts of the amplicons were loaded on a 1.5% agarose gel (lanes 2, 5 and 8: 2 µl, 5 µl and 20 µl respectively). The amplicons were then sequenced. To control that the amplification was not the result of DNA contamination, we did a second amplification with the same protocol, except that we avoid the reverse transcription step. Again, different amount of amplicons were loaded on a 1.5% agarose gel (lanes 3, 6 and 9 : 2 µl, 5 µl and 20 µl respectively). A molecular size marker was loaded on lanes 1, 4, 7 and 10 (Smart Ladder MW-1700-10 from Eurogentec, Seraing, Belgium).

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