PhiX174 DNA was completely digested by BsuRI, phenol extracted, ethanol precipitated and dissolved in 10mM Tris-HCl (pH 7.6), 1mM EDTA. The marker yields the following 11 discrete fragments (in base pairs): 1353, 1078, 872, 603, 310, 281, 271, 234, 194, 118, 72.
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, 60mM EDTA, 60% glycerol.
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.
Recommendations for Use:
I. Loading on Agarose Gel
1. Prepare DNA Marker before
loading:
1ul (0.5ug) of D3930-35 DNA Marker
1ul of L3350 6X Loading Dye Solution
4ul of deionized water
2. Vortex gently just prior to use. Do not heat before loading.
3. Apply the prepared amount (6ul) of
the DNA Marker on a 5mm lane
of agarose gel.
4. Following electrophoretic separation
on gel, visualize the DNA bands by
ethidium bromide staining.
Use 0.1ug (0.2ul) of the DNA Marker (before dilution) per 1mm of an agarose gel lane width.
II. Loading on Polyacrylamide Gel
1. Prepare DNA Marker before
loading:
2ul (1ug) of D3930-35 DNA Marker
0.5ul of L3350 6X Loading Dye Solution
0.5ul of deionized water
2. Vortex gently just prior to use. Do not heat before loading.
3. Apply the prepared amount (3ul) of
the DNA Marker on a 5mm lane
of polyacrylamide gel.
4. Following electrophoretic separation
on gel, visualize the DNA bands by
ethidium bromide staining.
Use 0.2ug (0.4ul) of the DNA Marker (before dilution) per 1mm of a Polyacrylamide gel lane width. 310, 281, 271 bp bands migrate anomalously.
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