Preparation of Nucleic Acid Probes

Authored by: Leland J. Cseke , Rogelio Luis Cruz-Vera

Handbook of Molecular and Cellular Methods in Biology and Medicine

Print publication date:  December  2011
Online publication date:  April  2016

Print ISBN: 9781420069389
eBook ISBN: 9781439881958
Adobe ISBN:

10.1201/b11351-6

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Abstract

Molecular biology studies usually involve the analysis of gene expression and structural characterization of DNA and RNA molecules. These procedures rely heavily on nucleic acid hybridization, such as Southern blots, northern blots, dot blots, microarrays, in situ hybridization, DNA/RNA protection assays, nucleotide sequencing, and chemical/enzymatic structural mapping/footprinting, among others. One of the most important aspects of these protocols is the use of either radioactively or nonradioactively labeled nucleic acids (DNA or RNA) as probes that specifically hybridize with their complementary DNA or RNA strands. The quality of the probe plays an essential role in detecting specific DNA or RNA sequences of interest. Therefore, the preparation of a probe with high specific activity is critical in nucleic acid hybridization. 1 The present chapter describes in detail reliable methods for the labeling of DNA and RNA. 2 6 These methods are well established and have been routinely used in our laboratory.

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