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

Chapters

  • 00:04Title
  • 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

자동 번역

자동 번역

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

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

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