Materials for electrophoresis

ELECTROPHORESIS OF DNA AND RNA FRAGMENTS REQUIRES HIGH QUALITY REAGENS DEVOID OF ANY DNASE and RNASE ACTIVITY

PCR loading buffers are solutions of high density, which simplify loading of the samples into electrophoretic gels. Loading buffers contain dyes which migrate during electrophoresis in agarose gel together with DNA. Bromophenol blue and xylene cyanol are widely used dyes in loading buffers. Bromophenol blue and xylene cyanol migrate in agarose gel in 0.5x TBE buffer similarly to double stranded DNA of about 300 bps and 4000 bps, respectively. This relative mobility is not affected by agarose gel concentration at a range 0.5 - 1.4%.

Presence of the dyes could, however affect visibility and quantification of DNA fragments in gels stained with ethidium bromide. Therefore, loading buffer containing bromophenol blue alone was also introduced (loading buffer Edge). Size of many PCR products is about 300 bps and therefore other dyes are also used. Yellow load loading buffer includes orange G, which migrates at position of individual nucleotides. Even faster migration exhibits a new dye in Read Load. The same red dye is also present in some PCR Master Mixes or PCR Purple Taq DNA polymerase and therefore this loading buffer is useful alternative in such studies.

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PCR vkládací pufr Red load

Loading buffers are solutions with high density, which facilitate loading of DNA- or RNA-containing solutions into the wells of agarose gel. Loading buffer Red contains the same dye, which is present in PPP master mixes or in Taq-Purple DNA polymerase and is therefore especially suitable for loading of DNA markers for PCR fragments amplified with these polymerases. For 10 µl of PCR product or other DNA-containing solution, 2-3 µl of loading buffer are added.

An integral component of PCR techniques is electrophoretic separation of DNA fragments in agarose gel. However, not every agarose is suitable for preparation of gels and electrophoretic separation. PCR agarose is the agarose with optimal properties for separation of DNA and also RNA molecules. This agarose has no RNase or DNase activity and is suitable for electrophoretic separation and transfer of DNA and RNA to membranes (blotting). Low value of electroendoosmosis (EEO) contributes to the increased electrophoretic mobility of RNA or DNA in the gel

PCR loading buffer Yellow load

Loading buffers are solutions with high density, which facilitate loading of DNA- or RNA-containing solutions into the wells of agarose gel. PCR loading buffer Yellow load contains yellow dye orange G, which migrates in the area of individual nucleotides. For 10 µl of PCR product or other DNA-containing solution, 2 - 3 µl of loading buffer are added.

PCR loading buffer

Loading buffers are solutions with high density, which facilitate loading of DNA- or RNA-containing solutions into the wells of agarose gel. PCR loading buffer contains two dyes, Bromophenol blue and Xylen cyanol, which allow monitoring of DNA fragments migration during electrophoresis in agarose gel. For 10 µl of PCR product or other DNA-containing solution, 2 - 3 µl of loading buffer are added.

PCR ethidium bromide

It is a filtered solution of ethidium bromide (10 mg/ml) in PCR H2O.

PCR loading buffer Edge

Loading buffers are the solutions with high density, which facilitate loading of of DNA- or RNA-solutions into the wells in agarose gel. PCR loading buffer Edge contains bromphenol blue and enables simple detection and densitometric quantification of DNA fragments in the area around 5 kb. For 10 µl of PCR product or other DNA-containing solution, 2-3 µl of loading buffer are added.