Lambda DNA was completely digested by HindIII, phenol extracted, ethanol precipitated and dissolved in 10mM Tris-HCl (pH 7.6), 1mM EDTA. The marker yields the following 8 discrete fragments (in base pairs): 23130*, 9416, 6557, 4361*, 2322, 2027, 564, 125.
*The cohesive ends of the 12nt cos site of bacteriophage lambda from fragments of 23130bp and 4361bp may anneal and form an additional band at 27491 bp. These fragments may be separated by heating at 65 degrees C for 5min and then cooling on ice for 3min.
6x Loading Dye Solution (L3350):
0.2% bromophenol blue, 0.2% xylene cyanol, 60% glycerol, 60mM EDTA.
Fragment (bp) % ng DNA/0.5ug
23130 47.7 238.4
9416 19.4 97.1
6557 13.5 67.6
4361 9.0 45.0
2322 4.8 23.9
2027 4.2 20.9
564 1.2 5.8
125 0.3 1.3
Note:
One vial (0.05mg) is sufficient for ~100 applications.
Use 0.1ug (0.2ul) of the DNA Marker (before dilution) per 1mm of agarose gel lane width.
Recommended Use:
Vortex gently prior to use.
Prepare DNA Marker as follows:
1ul (0.5ug) D3930-20
1ul of L3350
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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