Protocol for loading 1 kb Plus DNA Ladder for Safe Stains (NEB #N0559)
For GelRed / GelGreen Precast Gels: Load 5 µl (0.25 µg) per gel lane
For SYBR Safe Precast Gels: Load 10 µl (0.5 µg) per gel lane
Protocol and Usage Notes:
- Loading recommendation assume the stains are being used strictly following the usage recommendations set by the manufacturer.
- 1 kb Plus DNA Ladder for Safe Stains is stable for at least 6 months at 25°C. For long term storage, store at 4°C or -20°C. If stored at -20°C, mix well after thawing. If not well mixed, the load may float out of the wells.
- The 1 kb Plus DNA Ladder for Safe Stains is not intended for precise quantification of DNA mass but can be used for approximating the mass of DNA in comparably intense samples of similar size.
- The 1 kb Plus DNA Ladder for Safe Stains might be used on SYBR® Green I precast agarose gels but might require further dilution and optimization. We recommend using SYBR® Green I as a post-electrophoresis stain for best results.
- We recommend using SYBR® Gold as a post-electrophoresis stain only, as recommended by the dye’s manufacturer.
- Most gel stains run in the opposite direction from that of the migrating DNA. As a result, the bottom of the precast gel might have a lower concentration of stain, which might result in un-uniform staining of the bottom bands. How much of the gel will end up with a lower stain concentration is highly dependent on the migration conditions (gel percentage, voltage, buffer, run time, etc.). This effect can be counteracted by adding some gel stain to the running buffer.
- The 1 kb Plus DNA Ladder for Safe Stains is not intended for use on short mini-gels or fast electrophoresis systems. We recommend a minimum of 7 cm of gel length for proper and efficient separation. Best results occur with migration at 5 V/cm (distance between electrodes) for 60 to 90 minutes.