Chapter SixNucleic Acid Hybridization- Principles Appl.pptVIP

Chapter Six Nucleic Acid Hybridization: Principles Applications Preparation of nucleic acid probes: - DNA: from cell-based cloning or by PCR. Probe is double stranded. Labeling by DNA polymerase-based DNA strand synthesis. - RNA: by transcription from DNA cloned in an expression vector. Probe is single stranded. Labeling by “run-off” transcription. - Oligonucleotide: by chemical synthesis. Probe is single stranded. Labeling is by end labeling. 06_01.jpg DNA and RNA probes could be labeled in vitro by one of two methods: - Strand synthesis. By using DNA or RNA as a template to gen