| Autor: |
Alvarez ML; Diabetes, Cardiovascular, and Metabolic Diseases, Translational Genomics Research Institute, 445 Fifth Street, Phoenix, AZ, 85004, USA, lucrecia.alvarez@asu.edu., Doné SC |
| Jazyk: |
angličtina |
| Zdroj: |
Methods in molecular biology (Clifton, N.J.) [Methods Mol Biol] 2014; Vol. 1182, pp. 321-59. |
| DOI: |
10.1007/978-1-4939-1062-5_27 |
| Abstrakt: |
Quantitative PCR arrays are the most reliable and accurate tool for analyzing the expression of a focused panel of genes relevant to a pathway or a disease state. PCR arrays allow gene expression analysis with the sensitivity, dynamic range, and specificity of a real-time PCR as well as the multi-gene profiling capability of a microarray. Differences among real-time PCR kits used in PCR arrays are largely restricted to the DNA polymerases and the detection methods used. In this chapter, we provide a step-by-step protocol for the two detection methods most commonly used in PCR arrays, known as SYBR(®) Green and TaqMan(®), which are based on two different approaches to detect PCR products. While SYBR(®) Green uses a binding dye that intercalates nonspecifically into double-stranded DNA, the TaqMan(®) approach relies on a fluorogenic oligonucleotide probe that binds only the DNA sequence between the two PCR primers. Therefore, only specific PCR product can generate a fluorescent signal in TaqMan(®) PCR. Here we also provide a comparison of the SYBR(®) Green and TaqMan(®) approaches and highlight their advantages and disadvantages to help the user to choose the best platform. |
| Databáze: |
MEDLINE |
| Externí odkaz: |
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