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Membranes for Northern and Southern Blotting

Effective separation and detection of RNA and DNA in molecular biology research is essential for gene discovery, sequencing, and mapping used in diagnostics and industrial applications. Northern and Southern blotting are standard molecular biology techniques for RNA and DNA identification and quantification, respectively.

Alfa Chemistry strives to simplify detection through consistent, easy-to-read Northern and Southern blot transfer. We provide highly sensitive transfer membranes for Northern and Southern blot detection methods. It can identify and quantify RNA and DNA even in low-abundance samples.

Clarity of Visual Results

Northern blotting is used to detect specific RNA sequences in a variety of RNA mixtures, which involves complex separation and hybridization procedures that result in the binding of labeled probes to the RNA sequence of interest for detection and visualization. Southern blotting is used to detect specific DNA sequences in large and complex DNA samples. It can also be used to determine the molecular weight of restriction fragments and measure the relative amount of DNA in different samples.

Steps involved in DNA/RNA blotting procedureFigure.1 Steps involved in DNA/RNA blotting procedure

Northern and Southern blotting analysis methods are based on the immobilization of nucleic acids. RNA and fragments are separated by size on the membrane to allow the use of macromolecular probes for identification, detection, and visualization. To obtain clear results, the transfer film used needs to have:

  • High sensitivity
  • Surface consistency
  • Correct sensitivity level
  • Low background noise

When the abundance of RNA and DNA molecules is low, they may be difficult to separate and visualize. Membrane and detection schemes must provide the sensitivity to detect small amounts of molecules of interest without background interference. Alfa Chemistry provides transfer membranes with high sensitivity, low background and batch-to-batch consistency for all radioactive and non-radioactive detection methods.

The Right Membrane for Your Application

The nylon transfer membrane series provides a series of chemical substances with a variety of adsorption characteristics, suitable for different applications. These membranes are hydrophilic in nature, so it is easy to wet the entire membrane. Their stability and durability ensure that they will not rupture, shrink, or tear during multiple cycles of hybridization, peeling, and reprobing.

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A membrane zeta potential can be adjusted by pH value, which is very suitable for a single probe or multiple rehybridization. Membrane B is a positively charged nylon membrane with the highest sensitivity for nucleic acid applications. Plus membrane has a very high isoelectric point, which means it will maintain a good charge as the pH changes. C membrane is suitable for professional applications because it can be derivatized by the coupling reaction of carboxyl groups on the pore surface.

Please refer to the following selection guide for more information.

Northern and Southern Transfer Membrane Selection Guide
Membrane AMembrane B/PlusMembrane C
DescriptionAmphoteric Nylon 6,6Positively-charged Nylon 6,6Negatively-charged Nylon 6,6
AdvantagesHigh sensitivity
Low background
Ability to strip and reprobe
Net change can be controlled by changing pH
Positive charge over broad pH range
Ability to strip and reprobe
Highest sensitivity for nucleic acid applications (membrane B)
Ability to strip and reprobe
Negative charge over broad pH range
Surface carboxyl groups can be derivatized
Works suited forColony/Plaque Lifts
DNA and RNA Transfers
Gene Probe Assays,
DNA Fingerprinting,
Nucleic acid Dot/Slot Blots,
Replica Plating, ELISAs
Positive charge over broad pH range
Ability to strip and reprobe
Highest sensitivity for nucleic acid applications (membrane B)
Reverse Dot Blots
Protein Immobilization
Affinity Purification
ELISAs
Detection MethodsRadiolabeled Probes, Enzyme-antibody Conjugates
  • Chemiluminescent
  • Chromogenic
Radiolabeled Probes, Enzyme-antibody Conjugates
  • Chemiluminescent
  • Chromogenic
  • Chemifluorescent
Radiolabeled Probes, Enzyme-antibody Conjugates
  • Chromogenic
Binding InteractionHydrophobic and ElectrostaticHydrophobic and ElectrostaticHydrophobic and Electrostatic

Our products and services are for research use only and cannot be used for any clinical purpose.

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