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Epigenetics & Chromatin

Nucleosome Assembly Control DNA

control DNA for quick chromatin assembly

The Nucleosome Assembly Control DNA will prove to be a key component of your in vitro chromatin assembly reactions. The 187 bp double-stranded DNA can be directly added to your purified histones or histone extracts resulting in mononucleosome formation that can be easily visualized on a gel by mobility shift. The Nucleosome Assembly Control DNA can be used as a control to validate the proper assembly of the components of your in vitro chromatin assembly reactions. Alternatively, the Nucleosome Assembly Control DNA can be used as a template to monitor assembly kinetics in the presence or absence of chromatin interacting proteins or compounds.

 
Name Format Cat No. Price  
Nucleosome Assembly Control DNA 50 µg 53502 $120 Buy Now

High Affinity Binding to Histone Octamers

The Nucleosome Assembly Control DNA is a 187 bp double-stranded DNA fragment that has high affinity for histone octamers. The 187 bp double-stranded DNA was produced by PCR amplification from a plasmid containing the well-characterized 601 sequence (Lowary, 1998). The 601 sequence was originally isolated from a randomly generated synthetic DNA library in experiments that used the SELEX methodology in a selection strategy designed to identify DNA that bound to histone octamers with high affinity.

In Vitro Mononucleosome Assembly on Nucleosome Assembly Control DNA

The ability of the Nucleosome Assembly Control DNA to assemble into mononucleosomes is shown in Figure 1. After incubating the Nucleosome Assembly Control DNA with purified recombinant histone octamers, a high molecular weight band is seen on a gel. This band co-migrates with a highly purified control mononucleosome preparation.

Mononucleosome Assembly occurs when Nucleosome Assembly Control DNA is incubated with recombinant Histone Octamers
Figure 1: Mononucleosome Assembly Occurs when Histone Octamers are Incubated with Nucleosome Assembly Control DNA.

Recombinant histones (H2A, H2B, H3 and H4) were combined at equimolar ratios with polyglutamic acid to allow histone octomers to form. 1.4 µg of octamer was incubated with 0.7 µg Nucleosome Assembly Control DNA at room temperature for 1 hour. Samples were run on a 1% agarose gel and stained with ethidium bromide.

Lane 1: 100 bp DNA ladder.
Lane 2: Control mononucleosomes prepared by dialysis and purified.
Lane 3: Nucleosome Assembly Control DNA + histone octamers.
Lane 4: Nucleosome Assembly Control DNA + histone octamers and deproteinized with Proteinase K.

References

1. Lowery & Widom (1998) J Mol Biol 13, 19-42.

Contents & Storage

The Nucleosome Assembly Control DNA is a 187 base pair double-stranded DNA fragment. 50 µg of the DNA is supplied lyophilized.

The DNA is shipped at room temperature and should be stored at -20°C upon receipt. The DNA is guaranteed stable for 6 months from date of receipt when stored properly.