Sybr Green Agarose Gel Staining

Classical method for agarose and polyacrylamide gels.

Sybr green agarose gel staining.

One gel was visualized with sybr safe dna gel stain and blue light transillumination while the other gel was visualized with ethidium bromide and uv transillumination. Dsgreen an equivalent of sybr green i is a sensitive dsdna binding dye which can be used for routine dna detection in agarose and polyacrylamide gels. Seven equal amounts of the pcr product were electrophoresed on duplicate agarose gels. Sybr safe stain is supplied as a 10 000x concentrate in dmso which can be used just like a solution of ethidium bromide.

A qpcr grade reagent is also available. Gel soaking gel pre staining and sample pre staining. Unlike ethidium bromide dsgreen is highly selective towards double stranded dna much less harmful and offers better sensitivity. However there has been no systematic comparative analysis of the staining efficiencies of these dyes.

Although sybr gold or sybr green i have been used in the loading buffer as a dna stain safer than ethidium bromide for agarose gel electrophoresis electrophoretic mobility of dna is altered and. There are three variants of the staining protocol. In the present study sybr gold sybr green i goldview and eb were compared. Staining agarose gels with sybr green i does not interfere with the transfer of nucleic acids to membranes or subsequent hybridization in southern or northern blot analysis as long as 0 1 0 3 sds is included in prehybridization and hybridization buffers to remove the dye.

On the other hand sybr green ii is a highly sensitive stain for postelectrophoresis staining of rna and ssdna in agarose or polyacrylamide gels. Because sybr green i has greater sensitivity for dsdna it is especially useful for assays where the presence of contaminating rna or ssdna. Although both sybr gold and sybr green alter electrophoretic mobility and thus dna size estimates they are cost effective alternatives to eb. Sybr safe stain can be mixed into an agarose gel for staining during electrophoresis or the gel can be incubated in a solution of sybr safe stain following electrophoresis.

Dsgreen an analog of sybr green i is a fluorescent dye that binds specifically to double stranded dna. Sybr green i nucleic acid gel stain is one of the most sensitive stains available for detecting double stranded dna dsdna in agarose and polyacrylamide gels.