title: Application of reverse transcription-PCR and real-time PCR in nanotoxicity research.
abstract: Reverse transcription-polymerase chain reaction (RT-PCR) is a relatively simple and inexpensive technique to determine the expression level of target genes and is widely used in biomedical science research including nanotoxicology studies for semiquantitative analysis. Real-time PCR allows for the detection of PCR amplification in the exponential growth phase of the reaction and is much more quantitative than traditional RT-PCR. Although a number of kits and reagents for RT-PCR and real-time PCR are commercially available, the basic principles are the same. Here, we describe the procedures for total RNA isolation by using TRI Reagent, for reverse transcription (RT) by M-MLV reverse transcriptase, and for PCR by GoTaq(®) DNA Polymerase. And real-time PCR will be performed on an iQ5 multicolor real-time PCR detection system by using iQ™ SYBR Green Supermix.
authors.affiliation: Department of Environmental and Occupational Health Sciences, School of Public Health and Information Sciences, University of Louisville, Louisville, KY, USA.
authors.forename: Yiqun
authors.initials: Y
authors.lastname: Mo
authors.forename: Rong
authors.initials: R
authors.lastname: Wan
authors.forename: Qunwei
authors.initials: Q
authors.lastname: Zhang
mesh_terms: D000818:Animals; D006801:Humans; D053583:Interleukin-1beta / Q000235:genetics / Q000378:metabolism; D020778:Matrix Metalloproteinase 2 / Q000235:genetics / Q000378:metabolism; D009000:Monocytes / Q000187:drug effects / Q000201:enzymology; D053758:Nanoparticles / Q000633:toxicity*; D036103:Nanotechnology / Q000379:methods*; D012313:RNA / Q000302:isolation & purification; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D011817:Rabbits; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D020298:U937 Cells
chemical_list: D053583:Interleukin-1beta; D012333:RNA, Messenger; D012313:RNA; D020778:Matrix Metalloproteinase 2