Protocol for loading TriDye™ Ultra Low Range DNA Ladder (NEB #N0558)
For polyacrylamide gels, load 5 µl (0.5 µg) of TriDye™ Ultra Low Range DNA Ladder per gel lane.
For agarose gels, load 10 µl (1 µg) of TriDye™ Ultra Low Range DNA Ladder per gel lane
Protocol and Usage Notes:
- 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.
- If using wide combs (10 mm), we recommend loading 15 µl (1.5 µg) of TriDye Ultra Low Range DNA Ladder per gel lane.
- When using the DNA ladder on agarose gels, we recommend using ethidium bromide at 0.5 ug/ml in both the gel and the migration buffer for proper visualization of the smallest DNA fragments. We also recommend that you do not cast gels exceeding 5mm in thickness, as the smaller DNA fragments might diffuse more easily and may be harder to efficiently stain.
- We recommend using TBE buffer in the agarose gels and electrophoresis buffers, as TBE buffer allows for a better resolution of the smaller DNA fragments. Fragments below 15 bp might not separate efficiently when using TAE buffer.
- We do not recommend using GelRed® or SYBR® dyes as precast gel stains, as these dyes might interfere with the migration of the smallest DNA fragments and result in an abnormal band pattern. We recommend using these dyes as post-electrophoresis stains only. Optimization might be required for proper visualization of all DNA fragments. Intensity of some bands might vary.
- For optimal visualization of all the bands, we recommend exposing the gel to UV without any casting tray whenever possible. Even UV-transparent trays can obscure DNA fragments under UV exposure.