Lambda DNA was completely digested by both EcoRI and HindIII, phenol extracted, ethanol precipitated and dissolved in 10mM Tris-HCl (pH 7.6), 1mM EDTA. The marker yields the following 13 discrete fragments (in base pairs): 21226*, 5148, 4973, 4268, 3530*, 2027, 1904, 1584, 1375, 947, 831, 564, 125.
The cohesive ends of the 12 nt cos site of bacteriophage lambda from fragments of 21226 bp and 3530 bp (indicated*) may anneal and form an additional band at 24756 bp. These fragments may be separated by heating at 65 degrees C for 5min and then cooling on ice for 3min.
Prepare DNA Marker as follows:
1ul (0.5ug) DNA marker
1ul of 6X Loading Dye Solution
4ul ddH2O
1. Vortex gently prior to use
2. Heat 5 min. @ 65 degrees C, cool on ice 3 min.
3. Apply 6ul DNA marker per 5mm of agarose gel lane width.
4. Visualize bands with ethidium bromide staining.
Storage and Stability:
For long-term storage, aliquot and store at -20 degrees C. Aliquots are stable for at least 6 months at -20 degrees C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer.