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Transcriptomics: Home

Introduction

This guide is designed to assist you in your transcriptomics research endeavors.

Please, do not hesitate to contact me for questions or comments

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Genomics

TRANSCRIPTOMICS:  This another discipline in the OMIC field and aims to understand the complete set of RNA of an organism, in particular factors that lead to alterations in RNA expression.

Methods for transcriptomic studies

Northern blot:  This procedure is similar to the familiar procedure known as the Western blot for protein detection except we are detecting RNA.  RNA are separated by size using electrophoresis and detected using a probe complementary to the RNA of interest. 

DNA microarray:  This technique utilize wells or spots containing a DNA probe for the gene of interest.  RNA is isolated from cells or tissue and converted to cDNA.  The cDNA is labeled (different colors for control and treatment) and loaded onto the microarray wells. Through fluorometric procedures, the differential expression of RNA among two samples can be determined.

Quantitative polymerase chain reaction (qPCR):  This technique is used to quantify gene expression on the mRNA level in cells or tissue.  mRNA is converted to cDNA and gene expression is determined by the number of PCR amplification cycles it takes to reach a threshold copy number of cDNA.

RNA sequencing:  Similar to DNA sequencing, this method focuses on a broad view of cellular or tissue level RNA expression by using next generation sequencing.

Nanostring nCounter:  This technology uses a color coded barcode probes to detect nucleic acid expression (PCR without amplification?)

Nanostring nCounter:  This technology uses a color coded barcode probes to detect nucleic acid expression (PCR without amplification?)

Chromatin immunoprecipitation (ChIP) with our without sequencing:  ChIP is a way to determine whether a transcription factor binds to a certain geneof interest.  ChIP can be followed by qPCR to determine transcription factor to one or a small set of genes.  ChIP can also be followed by sequencing to determine the full gamut of binding activity for a particular transcription factor. 

Subject Guide

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Joe Wu
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