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Adapting 3′ Rapid Amplification of CDNA Ends to Map Transcripts in Cancer
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JoVE Journal Genética
Adapting 3′ Rapid Amplification of CDNA Ends to Map Transcripts in Cancer

Adapting 3′ Rapid Amplification of CDNA Ends to Map Transcripts in Cancer

DOI:
10.3791/57318-v

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09:38 min

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March 28, 2018

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1Department of Pharmaceutical Sciences, College of Pharmacy and Health Sciences,Butler University

Capítulos

  • 00:04Título
  • 01:01Cell Lysis and RNA Precipitation
  • 03:12cDNA Synthesis
  • 04:21Primer Search for Gene Transcript
  • 06:00Optimization and Verification of 3′ RACE to Map the 3′ UTR Using Two Different Enzymes
  • 07:46Results: Nested Forward Primer Search
  • 08:12Results: Successful Mapping of Cancer Transcripts
  • 09:03Conclusion

Summary

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Tadução automática

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.

Tags

Keywords: 3′ RACEMRNA TranscriptsCancerNested PCRTranscript SequencingPolyadenylation SignalStop Codon3′ UTRFusion GenesSplice VariantsRNA ExtractionPhenol-chloroformRNA PrecipitationRNA Resuspension

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