Journal
/
Genetics
/
Adapting 3′ Rapid Amplification of CDNA Ends to Map Transcripts in Cancer
JoVE Journal
Genetics
A subscription to JoVE is required to view this content.  Sign in or start your free trial.
JoVE Journal Genetics
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

•

09:38 min

•

March 28, 2018

•

,
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

Automatic Translation

Automatic Translation

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

Article

Embed

ADD TO PLAYLIST

Usage Statistics

Related Videos

check_url/57318?article_type=v

Merging Absolute and Relative Quantitative PCR Data to Quantify STAT3 Splice Variant Transcripts
check_url/57318?article_type=v

Engineering Artificial Factors to Specifically Manipulate Alternative Splicing in Human Cells
check_url/57318?article_type=v

Rapid Deletion Production in Fungi via Agrobacterium Mediated Transformation of OSCAR Deletion Constructs
check_url/57318?article_type=v

High Resolution Fluorescent In Situ Hybridization in Drosophila Embryos and Tissues Using Tyramide Signal Amplification
check_url/57318?article_type=v

CRISPR-Mediated Reorganization of Chromatin Loop Structure
check_url/57318?article_type=v

Amplification of Near Full-length HIV-1 Proviruses for Next-Generation Sequencing
check_url/57318?article_type=v

Reverse Transcription-Loop-mediated Isothermal Amplification (RT-LAMP) Assay for Zika Virus and Housekeeping Genes in Urine, Serum, and Mosquito Samples
check_url/57318?article_type=v

A Murine Cell Line Based Model of Chronic CDK9 Inhibition to Study Widespread Non-Genetic Transcriptional Elongation Defects (TEdeff) in Cancers
check_url/57318?article_type=v

Discrimintion and Mapping of the Primary and Processed Transcripts in Maize Mitochondrion Using a Circular RT-PCR-based Strategy
check_url/57318?article_type=v

Investigation of the Transcriptional Role of a RUNX1 Intronic Silencer by CRISPR/Cas9 Ribonucleoprotein in Acute Myeloid Leukemia Cells

Read Article