A non-radioactive method for small RNA detection by northern blotting
نویسندگان
چکیده
منابع مشابه
A rapid method of non radioactive Northern blot analysis.
The electrophoretic determination of RNA size by Northern blot analysis is an important procedure that has required to date the use of radiolabelled cDNA and cRNA probes. Here, we describe a new method which allows the size of an mRNA to be visualized by use of oligonucleotide probes directly labelled with alkaline phosphatase (AP). One advantage compared to radiolabelled probes is that AP-labe...
متن کاملA sensitive non-radioactive northern blot method to detect small RNAs
The continuing discoveries of potentially active small RNAs at an unprecedented rate using high-throughput sequencing have raised the need for methods that can reliably detect and quantitate the expression levels of small RNAs. Currently, northern blot is the most widely used method for validating small RNAs that are identified by methods such as high-throughput sequencing. We describe a new no...
متن کاملNorthern blotting of RNA denatured in glyoxal without buffer recirculation.
A rapid and easy procedure for preparing Northern blots is described. RNA is denatured with glyoxal in the presence of ethidium bromide and glycerol, then electrophoresed through agarose in a buffer that does not require recirculation. Without any additional washes, the RNA is vacuum-blotted to a nylon membrane in NaOH, which simultaneously removes the glyoxal adducts. All of these steps plus p...
متن کاملNorthern blotting analysis.
Northern blotting analysis is a classical method for analysis of the size and steady-state level of a specific RNA in a complex sample. In short, the RNA is size-fractionated by gel electrophoresis and transferred by blotting onto a membrane to which the RNA is covalently bound. Then, the membrane is analysed by hybridization to one or more specific probes that are labelled for subsequent detec...
متن کاملHighly sensitive northern hybridization using a rapid protocol for downward alkaline blotting of RNA.
A simple and fast RNA gel blot procedure is described that uses 50 mM NaOH to simultaneously transfer and fix RNA to a positively charged nylon membrane. The RNA is transferred in a downward direction, and transfer is routinely completed within 2.5 h. The resulting blots give increased sensitivity over existing methods without affecting RNA integrity and can be used in both radioactive and nonr...
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ژورنال
عنوان ژورنال: Rice
سال: 2014
ISSN: 1939-8425,1939-8433
DOI: 10.1186/s12284-014-0026-1