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تعداد آیتم قابل مشاهده باقیمانده : 3 مورد
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Comparison of methods to quantify gene expression

Comparison of methods to quantify gene expression
  Amount of starting RNA required Sensitivity of test Analytic requirements Information obtained from the test Number of targets Relative cost Advantages Disadvantages
RT-PCR Low High Low to moderate Relative gene expression Few to several hundred Low
  • Straightforward protocol
  • Higher sensitivity
  • Needs careful primer selection
RNA in-situ hybridization High Low to moderate Low Presence and spatial localization of RNA of interest Few Low
  • Spatial localization in a tissue section or cell
  • Low sensitivity
  • Qualitative, not quantitative
Microarrays (oligonucleotide arrays) Low to moderate Moderate to high Moderate to high Relative gene expression Thousands Moderate
  • Simultaneous detection of thousands of genes
  • Higher resolution than older methods
  • Analysis limited to known genes
  • Decreased sensitivity for lowly expressed transcripts
Custom microarrays Moderate Moderate to high Moderate to high Relative gene expression between two conditions Several hundred Moderate
  • Simultaneous detection of hundreds of genes
  • Direct comparison of two samples
  • Limited reproducibility
Transcriptome sequencing Low to moderate High Very high Digital counts of each RNA molecule (as measure of gene expression) with excellent dynamic range Unlimited High
  • Higher resolution than microarrays
  • Large dynamic range, can detect low abundance transcripts
  • Detects novel transcripts
  • Can identify polymorphisms within a sequence
  • High cost
  • Low throughput
  • Large storage space needed for data
  • Intensive computational analysis
Northern blot High Low Low Presence and size of RNA of interest Few Low
  • Straightforward
  • Risk of RNA degradation
  • Radioactivity use
  • Lower sensitivity
  • Higher starting quantity of RNA
Ribonuclease protection assay High Moderate Low Presence and size of RNA of interest Few Low
  • Higher specificity for the gene
  • Better sensitivity
  • Complex protocol
  • Radioactivity use
Refer to UpToDate for discussions of the clinical applications.
RNA: ribonucleic acid; RT-PCR: reverse-transcription polymerase chain reaction.
Graphic 76106 Version 3.0

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