Molecular Cloning Fourth Edition, A Laboratory Manual, by Michael R. Green and Joseph Sambrook

Table of Contents

Expand All | Contract All

Analysis of Small RNAs by Quantitative Reverse Transcription PCR

(Protocol summary only for purposes of this preview site)

Northern hybridization reveals the expression level and at same time the size(s) of a small RNA; it is the gold standard for small RNA validation and quantification. However, northern hybridization cannot detect low-abundance small RNAs, nor can it survey large numbers of small RNA species. In contrast, quantitative RT-PCR is more sensitive and can be adapted for high throughput (Fig.1). This protocol is adapted from Applied Biosystems TaqMan Small RNA Assays (Life Technologies Corporation 2011). TaqMan small RNA assays are designed to quantify RNAs 17200 nucleotides long, including siRNAs and miRNAs. TaqMan small RNA assays rely on a validated set of oligonucleotides comprising a stemlooped RT primer, a pair of PCR primers, and a TaqMan probe. Additional information on quantification of RNA by quantitative RT-PCR can be found in Chapter 9.


Previous   |   Next

Save 30% & Get Free Shipping!*

Save 30% at checkout on our website.
(Limited time special offer.) Molecular Cloning Fourth Edition, A Laboratory Manual, by Michael R. Green and Joseph Sambrook
Molecular Cloning Fourth Edition, A Laboratory Manual, by Michael R. Green and Joseph Sambrook

Search for information about other protocols included in the book:

Read What Others Are Saying About Molecular Cloning:

Tom Maniatis wins Lasker

'Not Quite a Book Prize'

Get alerts about special offers from Cold Spring Harbor Laboratory Press free to your Inbox

* Free shipping to individuals in U.S. and Canada only