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Analysis of the nad6 t-element

Analysis of the nad6 t-element. A. nad6. 3’. 5’. nad6 pre-mRNA. (-179). (+40). nad6 mRNA. (-17/-16). nad6 t-element. At5S-Mega.H. Atnad6-Endo.3’.H. 5S rRNA. 5S rRNA. nad6 t-element. Atnad6-Endo.3’.R. Atnad6-6. At5S-Mega.R. At5S-5. B. C. mt RNA. marker. 500/501/489 bp.

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Analysis of the nad6 t-element

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  1. Analysis of the nad6 t-element A nad6 3’ 5’ nad6 pre-mRNA (-179) (+40) nad6 mRNA (-17/-16) nad6 t-element At5S-Mega.H Atnad6-Endo.3’.H 5S rRNA 5S rRNA nad6 t-element Atnad6-Endo.3’.R Atnad6-6 At5S-Mega.R At5S-5

  2. B C mt RNA marker 500/501/489 bp 404 bp 331 bp 250/242 bp 190 bp I 147 bp 111/110 bp D

  3. E *A single adenosine at the ligation site could be either assigned to nad6 or 5S rRNA, so that the nad6 t-element either ends at position -16 or -15. If the 5S rRNA is assumed to end at position 361,062 as noted in the database entry (nc_001284) and as always found in the mapping experiments of the cox1 t-element, the adenosine is derived from 5S rRNA. Thus the 5’ end of the nad6 t-element is located at position -15. ** If the 5S rRNA 3’ end is located at position 361,062 (see above), there are three non-encoded As. Additionally, if the 5’ end of the nad6 t-element is located at position -15, four non-encoded As are present.

  4. F G mt RNA marker 500/501/489 bp 404 bp 331 bp 250/242 bp 190 bp II 147 bp 111/110 bp H

  5. K *A single nucleotide at the ligation site could be either assigned to nad6 or 5S rRNA. Thus the nad6 t-elements ends at position +27 or+28. Supplementary Figure 29. Analysis of the nad6 t-element. A modified CR-RT-PCR using endogenous 5S rRNA as adapter molecule was carried to determine the 5’ end (B-E) and the 3’ end (F-K) of the t-element downstream of the nad6 transcript (A). The site of the endonucleolytic cleavage of the pre-mRNA is marked by a pair of scissors. Primers indicated in tables (C) and (G), respectively, were used and the PCR products were separated by agarose gel electrophoresis (B and F). The products (marked by an arrow and detailed in tables (D) and (H)) were excised, cloned and analyzed by sequencing. The ends identified within the individual clones are listed in tables (E) and (K), respectively.

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