Loading buffer for protein, DNA and RNA

Details:

Loading buffer is a kind of loading buffer used to dissolve and add samples in the research experiments of protein, nucleic acid, DNA and RNA. Protein, DNA and RNA are different samples, and their loading buffers are also different.

Tris HCl (trimethylolaminomethane hydrochloric acid) or Tris glycine and EDTA salt (disodium ethylenediamine tetraacetate) are mainly used as buffer, and SDS, DTT, bromophenol blue, glycerol, etc. need to be added.

Difference of loading buffer of protein, DNA and RNA

SDS can combine with protein, so that protein SDS complex has a large number of negative charges. At this time, the charge of protein itself is completely covered by SDS, which eliminates the difference of charge of various proteins; SDS can also break the intramolecular and intermolecular hydrogen bonds and destroy the secondary and tertiary structures of protein molecules. DTT can break the disulfide bond between cysteine residues, destroy the protein structure and eliminate the differences between protein structures. Finally, for proteins (subunits) without charge and structural differences, the electrophoretic speed is only related to their molecular weight. Bromophenol blue is used as an indicator for electrophoresis, which can roughly indicate the end time of electrophoresis.

Composition of nucleic acid loading buffer

Nucleic acids include DNA and RNA. The loading buffer mainly contains Tris HCl, dye xylene blue FF, bromophenol blue (indicator), etc. The indicators bromophenol blue and xylene cyanogen FF play the role of indicating the process of electrophoresis, so that we can stop electrophoresis in time. The loading buffer of DNA and RNA can be basically universal, and their bases are basically the same. The dyes in the buffer can bind to their bases and run out of bands. However, if DNA loading buffer is used, it will be mixed with some RNases, which may affect the experimental results. RNases need to be disposed of before use.

It should be noted that although DNA and RNA loading buffers can sometimes be common, they cannot be common with protein buffers. Desheng is a manufacturer of diagnostic reagent raw materials, It can provide buffers related to protein and nucleic acid research.