Analysis of cytokine mRNA levels in interleukin-4-transgenic mice by quantitative polymerase chain reaction

1992 ◽  
Vol 22 (5) ◽  
pp. 1179-1184 ◽  
Author(s):  
Cornelia Platzer ◽  
Günther Richter ◽  
Klaus Überla ◽  
Werner Müller ◽  
Helmut Blöcker ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Transgenic Mice ◽  
Quantitative Polymerase Chain Reaction ◽  
Interleukin 4 ◽  
Mrna Levels ◽  
Chain Reaction ◽  
Cytokine Mrna ◽  
Polymerase Chain

Microarray analysis of cumulus cells in women with ovarian endometriosis undergoing intracytoplasmic sperm injection

2020 ◽  
Vol 12 (2) ◽  
pp. 86-93
Author(s):  
Abdullah Karaer ◽  
Gorkem Tuncay ◽  
Berat Dogan ◽  
Nihan Tecellioglu ◽  
Yilmaz Cigremis
Keyword(s):  
Body Mass Index ◽  
Polymerase Chain Reaction ◽  
Quantitative Polymerase Chain Reaction ◽  
Cumulus Cells ◽  
Polymerase Chain Reaction Analysis ◽  
P Value ◽  
Mrna Levels ◽  
Chain Reaction ◽  
Ovarian Endometriosis ◽  
Polymerase Chain

Objective: The aim of this study was to find the significantly altered genes in cumulus cells of women with ovarian endometriosis by using microarray and quantitative polymerase chain reaction analysis. Methods: Thirty women with ovarian endometriosis and 30 age–body mass index matched controls (women with infertility as a result of pure male factor) were enrolled in this study. Cumulus cells from study participants who underwent controlled ovarian hyperstimulation were isolated mechanically. Microarray comparative genomic hybridization was used to compare the transcriptome of cumulus cells from women with ovarian endometriosis and controls. According to the different expression levels in the microarrays and their putative functions, KRAS, ZNF322, and SDHA were selected and analyzed by real-time quantitative polymerase chain reaction. Results: There was no significant difference in the basal conditions between patients with endometriosis and controls, such as age, body mass index, basal follicle stimulating hormone and estradiol levels, and total gonadotrophin dosage. The gene expression profile of cumulus cells from patients with endometriosis was significantly different from that of controls. A total of 295 genes were significantly up- or down-regulated (p-value < 0.05 and absolute fold change > 1.5). For all of the genes adjusted p-value was found to be 0.999. Polymerase chain reaction analysis showed that KRAS and ZNF322 mRNA levels in the cumulus cells of patients with ovarian endometriosis were significantly up-regulated compared to controls (fold changes: 3.05 and 3.22, respectively). Conclusion: KRAS and ZNF322 mRNA levels in the cumulus cells of patients with ovarian endometriosis were significantly up-regulated.

AN IMPROVED METHOD FOR INHIBITOR FREE NUCLEIC ACIDS FROM POLYPHENOL RICH LEAVES OF MUSA SPP

2017 ◽  
Vol 23 (1) ◽  
Author(s):  
N.NANDHA KUMAR ◽  
K. SOURIANATHA SUNDARAM ◽  
D. SUDHAKAR ◽  
K.K. KUMAR
Keyword(s):  
Polymerase Chain Reaction ◽  
Quantitative Polymerase Chain Reaction ◽  
Proteinase K ◽  
Chain Reaction ◽  
Banana Plant ◽  
High Molecular Weight Dna ◽  
Musa Spp ◽  
Leaf Tissues ◽  
Polymerase Chain

Excessive presence of polysaccharides, polyphenol and secondary metabolites in banana plant affects the quality of DNA and it leads to difficult in isolating good quality of DNA. An optimized modified CTAB protocol for the isolation of high quality and quantity of DNA obtained from banana leaf tissues has been developed. In this protocol a slight increased salt (NaCl) concentration (2.0M) was used in the extraction buffer. Polyvinylpyrrolidone (PVP) and Octanol were used for the removal of polyphenols and polymerase chain reaction (PCR) inhibitors. Proteins like various enzymes were degraded by Proteinase K and removed by centrifugation from plant extract during the isolation process resulting in pure genomic DNA, ready to use in downstream applications including PCR, quantitative polymerase chain reaction (qPCR), ligation, restriction and sequencing. This protocol yielded a high molecular weight DNA isolated from polyphenols rich leaves of Musa spp which was free from contamination and colour. The average yields of total DNA from leaf ranged from 917.4 to 1860.9 ng/ìL. This modified CTAB protocol reported here is less time consuming 4-5h, reproducible and can be used for a broad spectrum of plant species which have polyphenol and polysaccharide compounds.

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