Monitoring MicroRNA Expression During Embryonic Stem-Cell Differentiation Using Quantitative Real-Time PCR (qRT-PCR)
Quantitative real-time PCR (qRT-PCR) is a reliable method to determine and monitor microRNA (miRNA) expression profiles in different cells, tissues, and organisms. Although there are several different strategies for performing qRT-PCR to determine miRNA expression, all of them have two steps in common: reverse transcription for obtaining cDNA from mature miRNA sequence and standard real-time PCR for amplification of cDNA. This chapter demonstrates the application of TaqMan-based real-time PCR for determining miRNA expression profiles during mouse embryonic stem-cell differentiation. In this method, a mature miRNA sequence is first reverse transcribed into a long cDNA with a 40- to 50-nt miRNA-specific stem-loop primer; then, a standard real-time PCR reaction is performed for determining miRNA expression using a forward miRNA-specific primer, a universal reverse primer, and FAM dye-labeled TaqMan probes.
- Selective Transport of Cationized Fluorescent Topoisomerase into Nuclei of Live Cells for DNA Damage Studies
- Tissue Culture Methods
- Use of Liposomes Containing Carbohydrates for Production of Monoclonal Antibodies
- Xenogeneic Lung Transplantation Models
- 扫描隧道显微镜(scanning tunneling microscope,STM)
- Nonradioactive Methods for Detecting Activation of Ras-Related Small G Proteins
- Post-transplant Monitoring of Chimerism by Lineage-Specific Analysis
- Oligonucleotide-Targeted RNase H Protection Analysis of RNA-Protein Complexes
- 光系统Ⅱ(photosystem Ⅱ complex, PSⅡcomplex)
- 核质蛋白(nucleoplasmin)