scholarly journals Gene Expression Analysis of the Effect of Ischemic Infarction in Whole Blood

2017 ◽  
Vol 18 (11) ◽  
pp. 2335 ◽  
Author(s):  
Ayako Takuma ◽  
Arata Abe ◽  
Yoshikazu Saito ◽  
Chikako Nito ◽  
Masayuki Ueda ◽  
...  
Keyword(s):  
Gene Expression ◽  
Expression Analysis ◽  
Whole Blood ◽  
Gene Expression Analysis ◽  
Ischemic Infarction

scholarly journals Gene expression analysis of whole blood RNA from pigs infected with low and high pathogenic African swine fever viruses

2017 ◽  
Vol 7 (1) ◽  
Author(s):  
Crystal Jaing ◽  
Raymond R. R. Rowland ◽  
Jonathan E. Allen ◽  
Andrea Certoma ◽  
James B. Thissen ◽  
...  
Keyword(s):  
Gene Expression ◽  
Expression Analysis ◽  
Whole Blood ◽  
Gene Expression Analysis ◽  
African Swine Fever ◽  
High Pathogenic

scholarly journals Sensitive and Quantitative Measurement of Gene Expression Directly from a Small Amount of Whole Blood

2006 ◽  
Vol 52 (7) ◽  
pp. 1294-1302 ◽  
Author(s):  
Zhi Zheng ◽  
Yuling Luo ◽  
Gary K McMaster
Keyword(s):  
Gene Expression ◽  
Expression Analysis ◽  
Reverse Transcription ◽  
Whole Blood ◽  
Quantitative Measurement ◽  
Gene Expression Analysis ◽  
Rna Isolation ◽  
Multiplex Assay ◽  
Quality Data ◽  
Branched Dna

Abstract Background: Accurate and precise quantification of mRNA in whole blood is made difficult by gene expression changes during blood processing, and by variations and biases introduced by sample preparations. We sought to develop a quantitative whole-blood mRNA assay that eliminates blood purification, RNA isolation, reverse transcription, and target amplification while providing high-quality data in an easy assay format. Methods: We performed single- and multiplex gene expression analysis with multiple hybridization probes to capture mRNA directly from blood lysate and used branched DNA to amplify the signal. The 96-well plate singleplex assay uses chemiluminescence detection, and the multiplex assay combines Luminex-encoded beads with fluorescent detection. Results: The single- and multiplex assays could quantitatively measure as few as 6000 and 24 000 mRNA target molecules (0.01 and 0.04 amoles), respectively, in up to 25 μL of whole blood. Both formats had CVs <10% and dynamic ranges of 3–4 logs. Assay sensitivities allowed quantitative measurement of gene expression in the minority of cells in whole blood. The signals from whole-blood lysate correlated well with signals from purified RNA of the same sample, and absolute mRNA quantification results from the assay were similar to those obtained by quantitative reverse transcription-PCR. Both single- and multiplex assay formats were compatible with common anticoagulants and PAXgene-treated samples; however, PAXgene preparations induced expression of known antiapoptotic genes in whole blood. Conclusions: Both the singleplex and the multiplex branched DNA assays can quantitatively measure mRNA expression directly from small volumes of whole blood. The assay offers an alternative to current technologies that depend on RNA isolation and is amenable to high-throughput gene expression analysis of whole blood.

scholarly journals Comparison of phenol-based and alternative RNA isolation methods for gene expression analyses

2010 ◽  
Vol 75 (8) ◽  
pp. 1053-1061 ◽  
Author(s):  
Ksenija Jakovljevic ◽  
Milena Spasic ◽  
Emina Malisic ◽  
Jelena Dobricic ◽  
Ana Krivokuca ◽  
...  
Keyword(s):  
Gene Expression ◽  
Expression Analysis ◽  
Whole Blood ◽  
Gene Expression Analysis ◽  
Expression Patterns ◽  
Rna Extraction ◽  
Rna Isolation ◽  
Specific Gene ◽  
Isolation Methods ◽  
Gene Expression Analyses

The widespread use of gene expression analyses has been limited by the lack of a critical evaluation of the methods used to extract nucleic acids from human tissues. For evaluating gene expression patterns in whole blood or leukocytes, the method of RNA isolation needs to be considered as a critical variable in the design of the experiment. Quantitative real-time PCR (qPCR) is widely used for the quantification of gene expression in today?s clinical practice. Blood samples as a preferred RNA source for qPCR should be carefully handled and prepared to not inhibit gene expression analyses. The present study was designed to compare the frequently used guanidine thiocyanate-phenol-chloroformbased method (TRI Reagent?) with two alternative RNA isolation methods (6100 PrepStation and QIAamp?) from whole blood or leukocytes for the purpose of gene expression analysis in chronic myeloid leukemia (CML) patients. Based on the results of this study, for the best combination of yield and RNA extraction purity, taking into account the necessary amount of the clinical sample and performance time, the protocol using phenol-based TRI Reagent? for RNA extraction from leukocytes is suggested as the most suitable protocol for this specific gene expression analysis.

Optimizing RNA extraction yield from whole blood for microarray gene expression analysis

2004 ◽  
Vol 37 (9) ◽  
pp. 741-744 ◽  
Author(s):  
Jian Wang ◽  
John F. Robinson ◽  
Hafiz M.R. Khan ◽  
David E. Carter ◽  
James McKinney ◽  
...  
Keyword(s):  
Gene Expression ◽  
Expression Analysis ◽  
Whole Blood ◽  
Gene Expression Analysis ◽  
Rna Extraction ◽  
Extraction Yield ◽  
Microarray Gene Expression ◽  
Microarray Gene ◽  
Microarray Gene Expression Analysis
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