Lambda DNA was completely digested by EcoRI, phenol extracted, ethanol precipitated and dissolved in 10mM Tris-HCl, pH 7.6, 1mM EDTA. The marker yields the following 6 discrete fragments (in base pairs): 21226*, 7421, 5804, 5643, 4878, 3530*.
*The cohesive ends of the 12 nt cos site of bacteriophage lambda from fragments of 21226 bp and 3530 bp may anneal and form an additional band at 24756 bp. These fragments may be separated by heating at 65 degrees C for 5 minutes and then cooling on ice for 3 minutes.
6x Loading Dye Solution (L3350):
Supplied as a liquid in 10mM Tris-HCl, pH 7.6, 0.03% bromophenol blue, 0.03% xylene cyanol FF, 60% glycerol, 60mM EDTA.
Quantity of DNA in band:
Fragment % ng DNA
(bp)
21226 43.8 218.8
7421 15.3 76.5
5804 12.0 59.8
5643 11.6 58.2
4878 10.1 50.3
3530 7.3 36.4
Total 100 500
Note:
1.One vial is sufficient for ~100 applications.
2. Use 0.1ug of DNA marker (before dilution) per 1mm and an agarose gel lane.
Recommended Use:
Vortex gently prior to use.
Prepare DNA Marker as follows:
1ul (0.5ug) DNA marker
1ul of 6X Loading Dye Solution
4ul ddH2O
Heat for 5 minutes at 65 degrees C. Cool on ice for 3 minutes. Apply the prepared amount (6ul) of the DNA Marker on a 5mm lane of agarose gel. Following electrophoresis separartion on gel, visualize the DNA bands by ethidium bromide staining.
Storage and Stability:
May be stored at 4 degrees C for short-term only. Aliquot to avoid repeated freezing and thawing. Store at -20 degrees C. Aliquots are stable for 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.
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