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Related Concept Videos

Real Time RT-PCR02:57

Real Time RT-PCR

Real-time reverse transcription-polymerase chain reaction, or Real-time RT-PCR, is an analytical tool used to determine the expression level of target genes. The method involves converting mRNA to complementary DNA with the help of an enzyme known as reverse transcriptase, followed by the PCR amplification of the cDNA. These two processes can be performed simultaneously in a single tube or separately as a two-step reaction.
The real-time quantification of the number of amplified products is...
RACE - Rapid Amplification of cDNA Ends02:35

RACE - Rapid Amplification of cDNA Ends

Rapid Amplification of cDNA Ends, or RACE, is one of the most effective methods to obtain a full-length cDNA from an mRNA sequence between a known internal region to the unknown sequence at the 5’ or 3’ end. The unknown region is cloned in the cDNA by a gene-specific primer that binds the known end, and a hybrid primer that attaches a predefined anchor sequence to the unknown end of the cDNA. The sequence in between is amplified by PCR with an anchor primer and a gene-specific primer.
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Single Cell Multiplex Reverse Transcription Polymerase Chain Reaction After Patch-clamp
10:44

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Published on: June 20, 2018

Reverse-transcriptase polymerase chain reaction to detect extracellular mRNAs.

Sweta Rani1, Lorraine O'Driscoll

  • 1School of Pharmacy & Pharmaceutical Sciences, Panoz Institute, Trinity College Dublin, Ireland. srani@tcd.ie

Methods in Molecular Biology (Clifton, N.J.)
|September 8, 2011
PubMed
Summary

Extracellular messenger RNAs (mRNAs) from insulin-producing cells may serve as novel biomarkers. These circulating nucleic acids could potentially indicate beta cell mass and function, aiding in disease diagnosis.

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Last Updated: May 29, 2026

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Area of Science:

  • Biochemistry
  • Molecular Biology
  • Endocrinology

Background:

  • Extracellular nucleic acids in serum/plasma are potential diagnostic markers for diseases like cancer and diabetes.
  • Investigating extracellular messenger RNAs (mRNAs) from insulin-producing cells offers a novel diagnostic avenue.

Purpose of the Study:

  • To explore the correlation between extracellular mRNA levels and the mass/function of insulin-producing cells.
  • To determine if specific mRNAs can be used as biomarkers for beta cell assessment.

Main Methods:

  • Reverse transcription-polymerase chain reaction (RT-PCR) was employed to detect and quantify various mRNAs.
  • Analyses included Pdx1, Npy, Egr1, Pld1, Chgb, InsI, InsII, and Actb mRNA.
  • Experiments were conducted on MIN6, MIN6 B1, and Vero-PPI cells and their conditioned media in biological triplicates.

Main Results:

  • Reproducible amplification of target mRNAs was achieved from the tested cell lines and their conditioned media.
  • This indicates that beta cells actively transcribe and release these mRNAs into their extracellular environment.
  • The findings demonstrate the presence and detectability of specific extracellular mRNAs.

Conclusions:

  • Extracellular mRNAs secreted by insulin-producing cells are detectable.
  • These secreted mRNAs show potential as extracellular biomarkers for evaluating beta cell mass and function.
  • Further research could validate these findings for clinical diagnostic applications.