Optimization and Verification of 3' RACE to Map the 3' UTR Using Two Different Enzymes
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Results: Nested Forward Primer Search
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Results: Successful Mapping of Cancer Transcripts
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Conclusion
Transcrição
The overall goal of this procedure is to determine the sequences of mRNA transcripts from the three-prime end to regions within the protein coding region by using transcript-specific nested primers in two subsequent PCRs and sequencing the purifie
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The two different 3' rapid amplification of cDNA ends (3' RACE) protocols described here make use of two different DNA polymerases to map sequences that include a segment of the open reading frame (ORF), the stop codon, and the entire 3' UTR of a transcript using RNA obtained from different cancer cell lines.