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تعداد آیتم قابل مشاهده باقیمانده : -9 مورد

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.

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