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Analysis of ATP1 Transcript Ends by CR-RT-PCR in Different RNA Samples

This study utilizes CR-RT-PCR to analyze ATP1 transcript ends in total RNA, mtRNA, and marker samples of various sizes. The method involves primer sets, gel electrophoresis, sequencing, and clone sequencing to identify specific ends. The results reveal insights into UTR length discrepancies and precise end assignments.

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Analysis of ATP1 Transcript Ends by CR-RT-PCR in Different RNA Samples

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  1. CR-RT-PCR analysis of atp1 transcript ends A B total RNA mt RNA marker 2,000 bp 1,500 bp 1,000 bp 750 bp I II C III 500 bp 250 bp

  2. D § to §§§§: 1 to 4 nucleotides could be assigned either to the 5‘ or to the 3‘ end. So actual 5‘ and/or 3‘ UTRs are 1 to 4 nucleotides longer than listed here.

  3. E Supplementary Figure 1. CR-RT-PCR analysis of atp1 transcript ends. CR-RT-PCR was carried out with the primers indicated in table (C) and the products were separated by agarose gel electrophoresis (A). Prominent products (marked by an arrow) were excised and analyzed by sequencing. The ends identified within the individual PCR products are listed in table (B). The CR-RT-PCR approach used to look specifically for the -1,898 5’ end is outlined in (D) and (E). CR-RT-PCR was carried out with the primers listed in table (E) and mtRNA. The products of the second PCR amplification were cloned and 30 individual clones sequenced. The ends detected within these clones are listed in table (D).

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