MRNA polymerase tissue expression

Godfrey TE, Kim S-H, Chavira M, et al. Quantitative mRNA expression analysis from formalin-fixed, paraffin-embedded tissues using 5 nuclease quantitative reverse transcription-polymerase chain reaction. J. Mol. Diagn. 2000 2 84-91. 

Cytokine profiling has also been measured as a function of changes in cytokine mRNA expression using either reverse transcription polymerase chain reaction (RT-PCR) [87, 91-93] or ribonuclease protection assay (RPA) [94-97], Measurement of cytokine transcripts by RT-PCR revealed that prolonged exposure to TMA induced increased levels of IL-4 mRNA expression compared with treatment with DNCB [87,92-93]. However, expression of the type 1 cytokine IFN-y by DNCB-activated LNC was variable and failed to discriminate between contact and respiratory allergens [87,91,93). A similar profile was observed for freshly isolated tissue analyzed by RPA. This somewhat less... 

The potential contribution of cytokines to the chronic inflammatory process of asthma has been made possible mainly by looking for the presence of cytokines using immunohistochemical techniques on airway mucosal tissues obtained from the proximal airways of patients with asthma. In addition, localization of cytokine mRNA by in situ hybridization or its detection by polymerase chain reaction (PCR) has also been used, although expression of mRNA may not necessarily mean that the protein is produced. The exact contribution of individual cytokines can best be surmised from studies of their effect in cells in vitro or in animals, particularly with the use of blocking antibodies, although extrapolation to the situation in disease must be made with some caution. 

FIGURE 2 Expression of exon 5 1- and exon 5 2-containing NOSl mRNAs in human tissues. Reverse transcription was performed using random primers and RNA (2 /ig) derived from leukocyte-enriched platelets (lanes 1 and 2), leukocyte-depleted platelets (lanes 3 and 4), skeletal muscle (lanes 5 and 6), or cerebellum (lanes 7 and 8) with (lanes 1, 3, 5, and 7) or without (lanes 2, 4, 6, and 8) prior treatment with RNase A and T1 followed by polymerase chain reaction (PCR) using Taq DNA polymerase and one twentieth of the reaction mixture and primer pairs specific for either (A) exon 5 2 or (B) exon 5 1. The PCR products were electrophoresed using 3% NuSieve GTG agarose (FMC Bioproducts), transferred to a Duralon-UV membrane (Stratagene), and hybridized with P-labeled probes specific for either (A) exon 5 2 or (B) exon 5T. nt. Nucleotides. 

NIS mRNA such as by northern blotting. Also, through the process of reverse transcription and a polymerase chain reaction (hereafter referred to as RT-PCR), a DNA strand identical to the RNA strand is formed and detected in a very sensitive assay. These RNA-based assays are two methods that can determine whether the symporter mRNA was present in a given tissue. Detection of symporter mRNA would indicate that the gene was being expressed. 

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