ABSTRACT
Probes are small (15-30 bases long) nucleotide sequences used to detect the presence of complementary sequences in nucleic acid samples. This is achieved by permitting the probes to base pair
10.1 Introduction .......................................................................................................................... 197 10.1.1 RNA versus DNA Probes ......................................................................................... 197 10.1.2 Probes ....................................................................................................................... 197
10.1.2.1 Preparation of Probes ................................................................................. 198 10.2 Preparation of High Speci c Activity Radiolabeled RNA Probes ....................................... 198
10.2.1 Introduction .............................................................................................................. 198 10.2.2 Protocol ..................................................................................................................... 199
10.3 Preparation of Non-Radiolabeled RNA Probes ....................................................................200 10.3.1 Digoxigenin Labeling ...............................................................................................200 10.3.2 Biotin Labeling of Nucleic Acids ............................................................................. 201 10.3.3 Fluorescent Labeling of Nucleic Acids ..................................................................... 201
10.4 Preparation of RNA Probes by PCR, Using Ampli ed RNA Technology .......................... 201 10.4.1 Advantages of RNA Ampli cation........................................................................... 201 10.4.2 RNA Ampli cation Procedure for Microarrays ....................................................... 201 10.4.3 Protocol for aRNA Ampli cation ............................................................................202
10.5 Preparation of RNA Probes for In Situ Hybridization .........................................................205 10.5.1 Advantages of RNA Probes for In Situ Hybridization .............................................205 10.5.2 Preparation of RNA Probes by PCR-Based Approach .............................................206 10.5.3 Protocol for RNA Probe Synthesis from PCR Product ............................................206 10.5.4 Custom GeneDetect® One-Step RNA Probe Synthesis Templates
