Northern blot analysis
Our protocol is applicable to analyses of all kinds of endogenous and exogenous RNAs, falling within length ranges of 20–30 and 50–70 nt, corresponding to microRNA and pre-microRNA lengths, respectively. We have also investigated other steps of the procedure with the goal of enhancing the resolution of RNAs herein, we present several recommendations to do so. We have enhanced the resolution of the northern blot technique by optimizing the electrophoresis step. Earlier efforts to improve this method focused mainly on altering the sensitivity of short RNA detection. Northern blot analysis consists of resolving RNAs by gel electrophoresis, followed by transferring and fixing to nylon membranes as well as detecting by hybridization with radioactive probes.
Northern blot analysis how to#
137, 266–267 (1984).This protocol describes how to perform northern blot analyses to detect microRNAs and their precursors with single-nucleotide resolution, which is crucial for analyzing individual length variants and for evaluating relative quantities of unique microRNAs in cells. 'A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity'. A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity. Most of the available information that are know today about gene expression and RNA. Hybridization probe size control: optimized 'oligolabelling'. Northern blot technique is very useful in the analysis of RNA molecules. Isolation of biologically active ribonucleic acid from sources enriched in ribonuclease. Expression and regulation of the murine serum amyloid A (SAA) gene in extrahepatic sites. Overexpression of the epidermal growth factor receptor in human pancreatic cancer is associated with concomitant increases in the levels of epidermal growth factor and transforming growth factor alpha. This chapter details the method for analyzing RNA by Northern blotting, which basically involves the isolation of RNA, its size fractionation by electrophoresis. Enhanced expression of transforming growth factor beta isoforms in pancreatic cancer correlates with decreased survival. using methods confirmed by western blotting (WB) was lower than that of studies. Enhanced expression of the type II transforming growth factor beta receptor in human pancreatic cancer cells without alteration of type III receptor expression. The Northern Hemisphere residents have the highest Lyme borreliosis (LB. Meta-analysis of microarray data on pancreatic cancer defines a set of commonly dysregulated genes. Identification of disease-specific genes in chronic pancreatitis using DNA array technology. Microarray-based identification of differentially expressed growth- and metastasis-associated genes in pancreatic cancer. Northern blotting can be used to analyze a sample of. Cytotechnology 12, 289–314 (1993).įriess, H. A northern blot is a laboratory method used to detect specific RNA molecules among a mixture of RNA. The use of reverse transcriptase-polymerase chain reaction (RT-PCR) to investigate specific gene expression in multidrug-resistant cells. Pancreatic cancer: from bench to 5-year survival. Molecular pathogenesis of pancreatic cancer: advances and challenges. Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Method for detection of specific RNAs in agarose gels by transfer to diazobenzyloxymethyl-paper and hybridization with DNA probes.