So sánh southern blot và northern blot
Northern and Southern blotting are standard molecular biology techniques for identification and quantification of RNA and DNA, respectively. Effective isolation and detection of RNA and DNA in molecular biology research is critical to gene discovery, sequencing, and mapping used in diagnostics and industry applications. Show Crucial to the blotting process is the clarity of the visual resultsNorthern and Southern blot analysis methods are based on the immobilization of nucleic acid. Nucleic acid fragments are separated by size onto a membrane to allow use of macromolecular probes for identification, detection, and visualization. For unambiguous outcomes, to avoid re-work and delays, the transfer membrane used needs to have:
Northern blot transfers are used for the detection of specific RNA sequences among a mixture of diverse RNA. This technique can be used to characterize the expression of RNA from tissue samples or cell cultures. It involves a complex isolation and hybridization procedure which results in labeled probes bound to the RNA sequence of interest for detection and visualization. Southern blotting is a molecular biology technique used for the detection of a specific DNA sequence in large, complex samples of DNA. The Southern blot method may also be used to determine the molecular weight of restriction fragments and to measure relative amounts of DNA in different samples. DNA samples can be obtained from tissue or isolated cell cultures. Transfer membrane sensitivity can impact ease of detectionOne of the challenges faced while identifying RNA or DNA fragments in samples through Northern and Southern blot analysis, is the relative abundance of RNA and DNA in the sample. When RNA and DNA molecules are less abundant, they can be difficult to isolate and visualize. The membrane and detection protocol must provide the sensitivity to detect low numbers of molecules of interest without picking up background interference. A clear picture with low background is required to correctly quantify RNA expression patterns and overall DNA presence. Cytiva supplies transfer membranes that demonstrate high sensitivity, low background, and lot-to-lot consistency for all radioactive and nonradioactive detection methods. Northern blotting protocol
Southern blotting protocol
Choosing the right membrane for your applicationThe Biodyne™ Nylon Transfer Membrane range (Pall™ Life Sciences products) offers a selection of chemistries with versatile adsorption properties for different applications. Our Biodyne A, Biodyne B, and Biodyne Plus membranes offer high sensitivity, low background, and lot-to-lot consistency for all radioactive and non-radioactive detection methods. These membranes are intrinsically hydrophilic resulting in easy wetting across the membrane. Their stability and durability ensure that they will not crack, shrink, or tear when subjected to multiple cycles of hybridization, stripping, and reprobing. Biodyne A membrane: Amphoteric meaning the membrane zeta potential can be modulated by changes in pH. Well-suited for single probe or multiple rehybridizations, and applications where background is troublesome. Biodyne B Membrane: Positively charged membrane. Our highest sensitivity nylon membrane for nucleic acid applications. Biodyne Plus Membrane: Positively charged with an extremely high isoelectric point, which maintains its charge with changing pH. Suitable for chemifluorescent detection. With certain nonradioactive detection systems, it’s more sensitive than Biodyne A membrane while exhibiting lower background than Biodyne B membrane (refer to selection guide table in this article). Biodyne C Membrane: Part of the Biodyne range, this membrane is applicable for specialist applications because it can be derivatized by coupling reactions through the carboxyl groups on the pore surfaces. |