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Northern Blot


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Northern blot is a commonly used molecular biology approach for the evaluation of gene expression at transcriptional levels (1). Northern blot is also known as RNA blot, as it reflects gene expressions via detection of the mRNA levels of target genes. A routine Northern blotting procedure starts with total RNA extraction, which is followed by separation of RNAs by electrophoresis. The blotting step refers to the transferring of separated RNAs to a nylon membrane, which is then exposed to specific probes for RNA detection. The probes can be either DNA or RNA with complementary sequences to target RNAs, and are often tagged with radioactive isotopes or chemiluminescent group (Figure 1).


Figure 1. A schematic description of the Northern blot assay (from Wikipedia)

Compared to other protein expression assays, the Northern blot technique have the following advantages (2):

  • High sensitivity (especially when using radioactive probes)
  • High specificity
  • Allows visualization of RNA size
  • Allows detection of alternate splice products
  • Nylon membranes are easy to store and can be re-probed
  • Easy to perform

Lifeasible, as a leading plant biotechnology company, has years of experience in plant molecular assays, as well as well-established platforms with advanced experimental equipment and solid technical support. To save your time and effort, we proudly provide you with one-stop Northern blot services that covers the following aspects:

  • Probe design and construction
  • RNA isolation
  • RNA separation by denaturing gel
  • Gel transferring (blotting)
  • Probe hybridization
  • Probe detection
  • Result analyzing and reporting

Moreover, to cope with different research goals, and specific status of each project, we offer customized protocols with optimized experimental equipments, buffers and solutions. our team of skilled scientists and experts are always ready to contribute their intelligence and efforts to your research.

References

  1. Alwine JC, Kemp DJ, & Stark GR (1977) Method for detection of specific RNAs in agarose gels by transfer to diazobenzyloxymethyl-paper and hybridization with DNA probes. Proc Natl Acad Sci U S A 74(12):5350-5354.
  2. Streit S, Michalski CW, Erkan M, Kleeff J, & Friess H (2009) Northern blot analysis for detection and quantification of RNA in pancreatic cancer cells and tissues. Nat Protoc 4(1):37-43.
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