Blotting, Northern. Medical search. FAQ

agarose gel
In order to prepare the reverse northern membrane, cDNA sequences for transcripts of interest are immobilized on nylon membranes, which can be accomplished by use of dot blots or bidirectional agarose gel blotting and UV fixation of the DNA to the membranes. (wikipedia.org)
typically through transferring (blotting) RNA fragments from an agarose gel to a nitrocellulose filter followed by detection with a suitable probe. (thefreedictionary.com)
The freshly isolated mouse CD1 tissue total RNA samples (20ug each) are fractionated though large formaldehyde-denatured agarose gel eletrophoresis, blotted onto a positively charged nylon membranes, and immobilized by UV light. (zyagen.com)

experimentally confirmed
The expression of SraJ was experimentally confirmed by Northern blotting. (wikipedia.org)

Molecular Biology
The northern blot, or RNA blot, is a technique used in molecular biology research to study gene expression by detection of RNA (or isolated mRNA) in a sample. (wikipedia.org)
Molecular biology experience including isolation of RNA, DNA, and proteins, Western and Northern blot analysis , RTPCR analysis of mRNAs, and immunohistochemistry are desirable. (thefreedictionary.com)

procedure
It is a play on words from Southern Blot , an essentially identical procedure used to detect DNA developed by the English scientist Ed Southern . (everything2.com)
A general blotting procedure starts with extraction of total RNA from a homogenized tissue sample or from cells. (wikipedia.org)
It is mRNA which is isolated and hybridized in northern blots.The formaldehyde was use in electrophoresis gel as a denaturant because the sodium hydroxide treatment used in the Southern blot procedure would degrade the RNA. (wikibooks.org)

Fragments
Fragments of DNA and RNA molecules separated by gel electrophoresis are transferred to a nylon or nitrocellulose membrane in a process termed as Southern and Northern blotting, respectively. (sigmaaldrich.com)
It is a variant of the northern blot in which the nucleic acid immobilized on a membrane is a collection of isolated DNA fragments rather than RNA, and the probe is RNA extracted from a tissue and radioactively labelled. (wikipedia.org)
The separated RNA fragments are then blotted onto a special filter paper so that each RNA molecule retains its position relative to all the other molecules. (wikibooks.org)

probes
Since the gels are fragile and the probes are unable to enter the matrix, the RNA samples, now separated by size, are transferred to a nylon membrane through a capillary or vacuum blotting system. (wikipedia.org)
Probes for northern blotting are composed of nucleic acids with a complementary sequence to all or part of the RNA of interest, they can be DNA, RNA, or oligonucleotides with a minimum of 25 complementary bases to the target sequence. (wikipedia.org)
Prepared reverse northern blot membranes are pre-hybridized in Denhardt's solution with SSC buffer and labeled cDNA probes are denatured at 100 °C and added to the pre-hybridization solution. (wikipedia.org)
In comparison to the Northern blot, the reverse northern blot is able to probe a large number of transcripts at once with less specificity with regard to probes than is required for Northern blot. (wikipedia.org)
Northern blot is limited by its ability to only probe with one mRNA at a time, while q-PCR requires transcripts to be long enough to generate primers for the sequence and probes can be costly. (wikipedia.org)
DNA microarrays operate by similar procedures to those used in the reverse northern blot, consisting of many DNA probes hybridized to a solid glass, plastic or silicon substrate which is probed with labeled RNA or cDNA. (wikipedia.org)
The filter is then exposed to radioactive probes that hybridize to complementary sequences in the blot. (wikibooks.org)
By modifying DNA oligonucleotides with LNAs using a design, in which every third nucleotide position was substituted by LNA, we could use the probes in northern blot analysis employing standard end-labelling techniques and hybridization conditions. (pubmedcentralcanada.ca)
The sensitivity in detecting mature microRNAs by northern blots was increased by at least 10-fold compared to DNA probes, while simultaneously being highly specific, as demonstrated by the use of different single and double mismatched LNA probes. (pubmedcentralcanada.ca)
Besides being highly efficient as northern probes, the same LNA-modified oligonucleotide probes would also be useful for miRNA in situ hybridization and miRNA expression profiling by LNA oligonucleotide microarrays. (pubmedcentralcanada.ca)
Detection of RNA that has been electrophoretically separated and immobilized by blotting on nitrocellulose or other type of paper or nylon membrane followed by hybridization with labeled NUCLEIC ACID PROBES. (mcw.edu)
Images of tissue total RNA used for construction of Northern blots and analysis with cDNA probes. (zyagen.com)

mRNA
As we demonstrated previously by Northern blot analysis (4) and semiquantitative RT-PCR (28), expression of human ABCA1 mRNA in macrophages is regulated in a differentiation-dependent fashion. (thefreedictionary.com)
To determine if Roundup reduced StAR protein levels by decreasing StAR mRNA levels, we performed Northern blot analysis (Figure 7). (thefreedictionary.com)
The expression of MIP-3α messenger RNA (mRNA) was analyzed in human multiple tissue Northern blots (Fig. 2 A). MIP-3α mRNA was highly expressed in lung and in inflammed tissues such as tonsil and appendix. (nih.gov)

sRNAs
The transcription of sRNAs was confirmed by Northern blot. (wikipedia.org)
The sRNAs were characterised through RACE mapping and Northern blot experiments. (wikipedia.org)

tissues
Reverse Northern blot, much like the northern blot upon which it is based, is used to determine levels of gene expression in particular tissues. (wikipedia.org)
Second, the Northern blot analysis of RNA samples from many different tissues enables researchers to determine which specific tissue a gene is expressed in along with the relative levels of its expression in all cells where transcription is occurring. (wikibooks.org)
Ready-to-hybridize Northern blots of total RNA of tissues that are difficult to obtain because of size, high RNase content, and anatomical complexity which requires knowledge, experience, and technical skill to exactly dissect the area of interest freely of surrounding tissues. (zyagen.com)
Saving labor by eliminating tedious and time-consuming laboratory procedures (harvesting and homogenization of tissues, RNA extraction, RNA quantification, gel electrophoresis, and blotting). (zyagen.com)
Every step of preparation of blots, from harvesting tissues and extraction of RNA to the blotting, is carefully monitored to ensure the superior quality and performance. (zyagen.com)

membrane
The term 'northern blot' actually refers specifically to the capillary transfer of RNA from the electrophoresis gel to the blotting membrane. (wikipedia.org)
A nylon membrane with a positive charge is the most effective for use in northern blotting since the negatively charged nucleic acids have a high affinity for them. (wikipedia.org)

Polyacrylamide
Most miRNA researchers use northern blot analysis combined with polyacrylamide gels to examine the expression of both the mature and precursor miRNAs, since it allows both quantitation of the expression levels as well as miRNA size determination ( 5 , 7 , 8 , 13 ). (pubmedcentralcanada.ca)

analysis
The Northern blot is similar to the Southern blot except that RNA instead of DNA is the subject of analysis in this technique. (wikibooks.org)
Northern Blot is a derivative of the Southern blot , which, like the Southern Blot, utilizes electrophoresis separation analysis and a hybridization probe for detection. (wikibooks.org)
RNA purification is a critical first step for preparing biological samples for a multitude of analytical techniques including real-time quantitative PCR, reverse transcription PCR, microarrays, Northern blot analysis , nuclease protection assays, RNA mapping, in vitro translation and DNA library construction. (thefreedictionary.com)
In addition, based on the array and Northern blot analysis of miRNAs 161, 168a&b, 167a&b and 171a, we might speculate that they have a more involved role in the plants defense to viral infection as opposed to miRNAs 159 and 172a. (thefreedictionary.com)
For Northern blot analysis , l0[micro]g of total RNA of each sample was denatured by incubation for 1 h at 50 [degrees]C in a solution of 0. (thefreedictionary.com)
Northern blot analysis of total RNA from planktonic and biofilm-grown cells demonstrated that ALS gene expression is differentially regulated between the two growth forms, with additional genes expressed in biofilms (4,5). (thefreedictionary.com)
We have not quantified the response by DD RT-PCR and Northern blot analysis compared with the arrays because limited amounts of samples required the use of different RNA samples in these experiments. (thefreedictionary.com)
Finally, 10 of these pairs were further analyzed using northern blot analysis with strand-specific riboprobes. (thefreedictionary.com)
The sodium/iodide symporter outside the thyroid gland, however, does not appear to be affected by thyroidstimulating hormone, even though the sequence appears identical to that of the thyroid receptor on Northern blot analysis and DNA sequencing analysis. (thefreedictionary.com)
Powerful and continually evolving methods, such as short-read sequencing (RNA-seq), microarray analysis, quantitative real-time RT-PCR, as well as traditional methods for differential gene expression studies using multiplex endpoint PCR and northern blot analysis are employed by scientists to analyze gene function, identify new therapeutic and diagnostic targets, and to map pathways involved in development and disease. (thefreedictionary.com)
MEASUREMENTS AND MAIN RESULTS: Theaflavin inhibited tumor necrosis factor-alpha-mediated interleukin-8 gene expression, as measured by luciferase assay and Northern blot analysis , at concentrations of 10 and 30 microg/mL. (thefreedictionary.com)
We describe here a new method for highly efficient detection of microRNAs by northern blot analysis using LNA (locked nucleic acid)-modified oligonucleotides. (pubmedcentralcanada.ca)
Consequently, a large amount of total RNA per sample is required for northern analysis, which is not feasible when the cell or tissue source is limited. (pubmedcentralcanada.ca)
Using Northern Blot Analysis, Vitelli et al. (wikipedia.org)
Northern blot analysis also indicated this RNA undergoes specific cleavage processing. (wikipedia.org)
The stability and abundance of the shorter 3' transcript is confirmed in both Northern blotting and deep sequencing analysis. (wikipedia.org)
However, northern blot analysis of this 477bp sequence yielded no results. (wikipedia.org)

1977
The northern blot technique was developed in 1977 by James Alwine, David Kemp, and George Stark at Stanford University, with contributions from Gerhard Heinrich. (wikipedia.org)
In order to avoid confusion, the following abbreviations will be used consistently throughout this article: Since its introduction in 1977, northern blot had been used extensively for RNA quantification despite its shortcomings: (a) time-consuming technique, (b) requires a large quantity of RNA for detection, and (c) quantitatively inaccurate in the low abundance of RNA content. (wikipedia.org)

Detection
Northern blotting involves the use of electrophoresis to separate RNA samples by size, and detection with a hybridization probe complementary to part of or the entire target sequence. (wikipedia.org)
Compared to other RNA quantification methods, such as northern blot, qRT-PCR is considered to be the most powerful, sensitive, and quantitative assay for the detection of RNA levels. (wikipedia.org)
However, the discovery of reverse transcriptase during the study of viral replication of genetic material led to the development of RT-PCR, which has since displaced northern blot as the method of choice for RNA detection and quantification. (wikipedia.org)

ncRNA
The expression of novel ncRNA of interest was confirmed by northern blotting. (wikipedia.org)

probe
Incubate the blot with labeled probe. (sigmaaldrich.com)
The transfer buffer used for the blotting usually contains formamide because it lowers the annealing temperature of the probe-RNA interaction, thus eliminating the need for high temperatures, which could cause RNA degradation. (wikipedia.org)
Commonly cDNA is created with labelled primers for the RNA sequence of interest to act as the probe in the northern blot. (wikipedia.org)
A labelled probe specific for the RNA fragment in question is incubated with the blot. (wikibooks.org)
The blot is washed to remove non-specifically bount probe and then a development step allows visualization of the probe that is bound. (wikibooks.org)
The integrity of the blotted RNA is tested by beta-actin specific probe. (zyagen.com)

nucleic
The transfer of macromolecules such as nucleic acids and proteins to solid-phase membranous support is known as blotting. (sigmaaldrich.com)

membranes
Blots are made of high quality total RNA samples which are subjected to rigorous quality control procedures before fractionation through large denatured agarose gels and blotting onto positively charged nylon membranes. (zyagen.com)

expression
With northern blotting it is possible to observe cellular control over structure and function by determining the particular gene expression rates during differentiation and morphogenesis, as well as in abnormal or diseased conditions. (wikipedia.org)
The reverse northern blot is a method by which gene expression patterns may be analyzed by comparing isolated RNA molecules from a tester sample to samples in a control cDNA library. (wikipedia.org)
A reverse northern blot can be used to profile expression levels of particular sets of RNA sequences in a tissue or to determine presence of a particular RNA sequence in a sample. (wikipedia.org)
Although DNA Microarrays and newer next-generation techniques have generally supplanted reverse northern blotting, it is still utilized today and provides a relatively cheap and easy means of defining expression of large sets of genes. (wikipedia.org)
Therefore, reverse northern has been used as one means of confirming hits from DD-PCR, or sequences with altered expression levels. (wikipedia.org)
The Northern blot is useful for the study of gene expression in two ways. (wikibooks.org)
The Northern blot is a valuable method used by researchers in determining gene expression patterns. (wikibooks.org)
For example, many scientists researching Huntington disease or breast cancer are able to determine the expression patterns of the genes responsible for these diseases using blotting techniques. (wikibooks.org)
The main difference between the two techniques is that the northern Blot is performed to study gene expression by analyzing RNA instead of DNA. (wikibooks.org)
Expression of 11 of them was confirmed by Northern blot. (wikipedia.org)
U45 was originally cloned from HeLa cells and expression verified by northern blotting. (wikipedia.org)

tissue
If you are interested in Northern blot constructed with any mouse RNA tissue of any strain or required blots to be specifically prepared to meet your application, our custom service scientists can make it for you. (zyagen.com)

protein
There also exists a similar, but different method of detecting protein s called a Western Blot . (everything2.com)
The other blotting techniques emerged from this method have been termed as Northern (for RNA), Western (for proteins), Eastern (for post-translational protein modifications) and Southwestern (for DNA-protein interactions) blotting. (sigmaaldrich.com)

researchers
To determine what makes grey horses more susceptible to melanomas, researchers have used different techniques such as the Northern Blot technique and Real-Time PCR. (wikipedia.org)

sequences
Southern blotting was introduced by Edwin Southern in 1975 as a method to detect specific sequences of DNA in DNA samples. (sigmaaldrich.com)

gels
The gels can be stained with ethidium bromide (EtBr) and viewed under UV light to observe the quality and quantity of RNA before blotting. (wikipedia.org)

lane
b) Invitrogen Northern territory blots (20 μg total RNA/ lane). (nih.gov)

Reverse
Reverse northern blotting was used in a 2013 study in Gene in which the author identified a number of genes responsible for early cold-resistance response in the cold-hardy citrus fruit Poncirus trifoliata. (wikipedia.org)

technique
Northern blotting (inglise northern blot , northern blot technique ) on meetod RNA osade identifitseerimiseks vastavates proovides, kus agaroosgeelis lahku viidud RNA lõigud viiakse üle nt nitrotselluloospaberile ja üleviimise tulemusel saab RNA lõike uurida nt radioaktiivselt markeeritud geeniproovi kaudu . (wikipedia.org)
Northern blotting takes its name from its similarity to the first blotting technique, the Southern blot, named for biologist Edwin Southern. (wikipedia.org)
Northern blotting is a laboratory technique that produces similar information. (wikipedia.org)

Methods
Hi out there, Are there some methods that would allow to have southern and northern on the same blot? (bio.net)

techniques
While northern blot or q-PCR are often used to confirm results, both techniques have drawbacks. (wikipedia.org)
The Northern blot, also known as the RNA blot, is one of the blotting techniques used to transfer DNA and RNA onto a carrier for sorting and identification. (wikibooks.org)
The three blotting techniques including the Northern blot have proven to be important and positive advancements in science. (wikibooks.org)

similar
20X SSC ( 93017 ) may be used in place of SSPE in similar blotting protocols. (sigmaaldrich.com)

total
Blots are made of total RNA that is treated with RNase-free DNase to remove residual DNA. (zyagen.com)

method
A Northern Blot is a method by which RNA is identified. (everything2.com)

samples
To create controls for comparison in a northern blot, samples not displaying the gene product of interest can be used after determination by microarrays or RT-PCR. (wikipedia.org)

positive
Hi everyone, my boss ask me to have a good positive control for my northern blot. (protocol-online.org)

problems
Hallo, I am having problems with Northern blots. (bio.net)
Good morning everybody, I have some problems with Northern blot. (protocol-online.org)

However
However, the entire process is commonly referred to as northern blotting. (wikipedia.org)

provides
First, the position of bands on the blot provides a direct measure of RNA size. (wikibooks.org)

results
Hi, I'm new to microRNA.I want to confirm the deep sequencing results.qrt-pcr and northern blot ,which way is better? (protocol-online.org)

room
Blots are shipped in a sealed bag at room temperature and are good for 1 year if stored at 4 o C in the sealed bags. (zyagen.com)

major
The major difference is that RNA, rather than DNA, is analyzed in the northern blot. (wikipedia.org)