Preexisting Type 2 Immune Activation Protects against the Development of Sepsis
Year: 2017
Type: article
Abstract: achieve further improvements in RT-qPCR, whereas switching to different qPCR master mix solutions has a negative effect on Ct values (Table E1).Finally, we compared the original and improved protocols with regard to both RNA isolation and RT-qPCR.A 7.4-fold increase in RNA yield was achieved by using the optimized RNA isolation protocol (Figure 1E).Moreover, optimization of the qPCR protocol resulted in significantly lower Ct values of GAPDH amplicons of all sizes compared with the original protocol (Table E2).In conclusion, we have developed a combined sputum processing/RNA extraction/RT-qPCR protocol to maximize the quantity and quality of RNA obtained from sputum, and to enhance detection of transcripts by qPCR.Successful elimination of bacterial DNA was shown to be a critical step in the optimization process.Our results should facilitate future gene-expression studies utilizing sputum.
Institution: University of Chicago
Language: English
Subfield: Epidemiology
Field: Medicine
Domain: Health Sciences
Open Access status: green
Awards: Mendelian Disease - Asthma Comorbidity to Find Subgroup-Specific Asthma Genes, IRF4+ respiratory dendritic cells in type 2 inflammatory responses, Airway inflammation and HLA-G in asthma, Type 2 Immune Responses Protect Against the Morbidity and Mortality of Sepsis, Translational Research at The University of Chicago (UL1), Functional Genomics of IL-33 expression and asthma risk, Training of Junior Faculty for Careers in Omics of Lung Diseases, Research training in respiratory biology
OpenAlex ID: W2767042436