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RNA-seq03:21

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The t-test is a statistical method used to compare the sample mean with a population mean or compare two means from two data sets. The test statistic is calculated from the standard deviation, mean, and number of measurements in the data set at a selected confidence interval and then compared to a table of critical values at this confidence level. If the test statistic is smaller than the critical value, the null hypothesis is accepted. In this case, we state that the difference between the...
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The present-day mitochondrial and chloroplast genomes have retained some of the characteristics of their ancestral prokaryotes and also have acquired new attributes during their evolution within eukaryotic cells. Like prokaryotic genomes, mitochondrial and chloroplast genomes neither bind with histone-like proteins nor show complex packaging into chromosome-like structures, as observed in eukaryotes. Unlike mitotic cell divisions observed in eukaryotic cells, mitochondria and chloroplasts...
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Comparative evaluation of HTG and TempO Seq targeted transcriptome profiling methods.

Antonio Fernández-Serra1,2, Raquel López-Reig1,2, Ignacio Romero3

  • 1Laboratory of Molecular Biology, Valencian Institute of Oncology, Valencia, Spain.

Scientific Reports
|January 24, 2026
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Summary

TempO-Seq offers a viable alternative for targeted RNA sequencing in molecular oncology research. It shows comparable performance to the discontinued HTG EdgeSeq HTP platform for multi-gene biomarkers in FFPE samples.

Keywords:
Biomarkers in molecular oncologyEndometrial cancerFFPE samplesTargeted RNA-seq

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Area of Science:

  • Molecular biology
  • Translational oncology
  • Molecular oncology

Background:

  • Transcriptomic profiling is crucial for molecular oncology, requiring reliable gene expression measurements.
  • Targeted RNA sequencing offers advantages for formalin-fixed paraffin-embedded (FFPE) samples.
  • The discontinuation of the HTG EdgeSeq HTP platform created a need for alternative targeted transcriptome analysis methods for FFPE specimens.

Purpose of the Study:

  • To systematically compare the HTG EdgeSeq HTP platform with the TempO-Seq platform.
  • To assess TempO-Seq's suitability as an alternative for targeted transcriptome analysis in molecular oncology research using FFPE samples.

Main Methods:

  • Analysis of 21 FFPE endometrial cancer samples and three RNA reference controls.
  • Evaluation of concordance between platforms across individual transcripts and multi-gene signatures.
  • Systematic technical comparison of HTG EdgeSeq HTP and TempO-Seq platforms.

Main Results:

  • Single-gene measurements showed limited concordance between the two platforms.
  • Multi-gene biomarkers demonstrated substantially higher agreement between platforms.
  • Aggregation across multiple probes mitigated platform-specific effects on biomarker agreement.

Conclusions:

  • TempO-Seq provides comparable performance to HTG EdgeSeq HTP for multi-gene biomarkers.
  • TempO-Seq is a robust alternative for targeted transcriptomic profiling of FFPE samples in molecular oncology research.
  • The TempO-Seq platform can fill the technical gap left by the discontinuation of HTG EdgeSeq HTP.