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ZR-96 ChIP DNA Clean & Concentrator™

Rapid high throughput (96-well) recovery of ultra-pure DNA from chromatin immunoprecipitation (ChIP), cell lysates, Proteinase K digested samples, PCRs, and other enzymatic reactions.
Plate design permits DNA elution at high concentrations into minimal volumes (≥10 µl/well).
Omits the use of organic solvents and the need for ethanol precipitation.
Eluted DNA is well suited for use in PCR, Next-Gen sequencing (ChIP-Seq), microarrays, Southern blot analysis, and other molecular applications.

Product Size Catalog # Price Qty
ZR-96 ChIP DNA Clean & Concentrator™ 2 x 96 Preps D5206
$286.00
ZR-96 ChIP DNA Clean & Concentrator™ 4 x 96 Preps D5207
$456.00

About ZR-96 ChIP DNA Clean & Concentrator™

The ZR-96 ChIP DNA Clean & Concentrator™ (ZR-96 ChIP DCC™) provides a hassle-free method for the rapid, high throughput purification and concentration of high-quality DNA from any step in a "standard" ChIP protocol.  This includes samples that have: A) undergone reverse cross-linking and Proteinase K or RNase A digestion following mechanical/enzymatically-mediated DNA shearing or B) reverse cross-linked samples eluted from chromatin-antibody-bead complexes.  Additionally, this product may also be used to purify DNA from PCR and other enzymatic reactions.  DNA purified using the ZR-96 ChIP DCC™ is suitable for PCR, Next-Gen sequencing (ChIP-Seq), arrays, as well as other molecular applications.


DNA Recovery Typically, up to 5 µg total DNA (per well) can be eluted into as little as 10 µl of low salt DNA Elution Buffer or water. For DNA 50 bp to 10 kb, the recovery is 70-90%. For DNA 11 kb to 23 kb, the recovery is 50-70%.
DNA Size Limits From ~50 bp to 23 kb.
Sample Sources Any step in a standard ChIP protocol including:
a) Samples that have undergone reverse cross-linking and Proteinase K or RNase A digestion following mechanical or enzymatically-mediated DNA shearing.
b) Reverse cross-linked samples eluted from chromatin-antibody-bead complexes in TES, 0.1M NaHCO3 and 1% SDS, or other buffers containing up to 1% SDS.
Note: This kit can also be used for DNA purification from PCR, restriction digests, kinase, phosphatase and other enzymatic reactions.
DNA Purity High-quality, purified DNA is eluted with elution buffer or water and is especially well suited for PCR, Next-Gen sequencing (ChIP-Seq), microarrays, Southern blot analysis, and other molecular applications.
Product Detergent Tolerance ≤5% Triton X-100, ≤5% Tween-20, ≤5% Sarkosyl, ≤1% SDS.



Figure 1: ZR-96 ChIP DNA Clean & Concentrator™ procedure. The ZR-96 ChIP DCC™ employs a single buffer system that allows for efficient DNA adsorption to the matrix of the supplied Zymo-Spin™ I-96 Plate. The DNA is washed twice then eluted with a small volume of elution buffer or water.
       

Figure 2: Quantitative recovery of DNA from cell lysates. The ZR-96 ChIP DNA Clean & Concentrator™ was used to purify DNA from lysates. The amount of DNA recovered was proportional to the lysate volume. Ultra-pure DNA isolated from 50, 100, and 150 µl cell lysates was eluted with 10 µl elution buffer and the DNA concentrations were determined using UV-Vis spectrophotometery.


Figure 3: Yeast ChIP PCR Analysis. S.cerevisiaecultures were incubated at 30°C for 45 min. in YEP with or without galactose. Following cross-linking, cell lysis, and DNA shearing, ChIP was performed using an antibody specific for RNA polymerase II. Reverse cross-linking was followed by Proteinase K digestion and DNA purification using the ChIP DNA Clean and Concentrator™. PCR was performed using primers to GALregions and the products were subsequently analyzed by agarose gel electrophoresis.

Note: All centrifugation steps should be performed between 3,000 - 5,000 x g.

  1. Add 5 volumes of ChIP DNA Binding Buffer to each volume of DNA sample. Mix briefly by vortexing.

    Example:    Add 250 µl ChIP DNA Binding Buffer to 50 µl eluent in TES or 0.1M NaHCO3 containing 1% SDS buffers from chromatin-antibody-Protein A agarose-bead complexes followed by reverse cross-linking and Proteinase K digestion.
  2. Transfer sample mixtures to the wells of a Zymo-Spin™ I-96 Plate mounted on a Collection Plate.
  3. Centrifuge for 5 minutes until sample mixtures have been completely filtered.  Discard the flow-through.
  4. Add 300 µl DNA Wash Buffer to each well of the Zymo-Spin™ I-96 Plate.  Centrifuge for 5 minutes.  Repeat wash step, but centrifuge for 15 minutes. 
  5. Add ≥ 10 µl DNA Elution Buffer or water directly to the column matrix in each well.  Transfer the Zymo-Spin™ I-96 Plate onto an Elution Plate and centrifuge for 5 minutes to elute the DNA.

    Ultra-pure DNA is now ready for use in PCR, Next-Gen sequencing (ChIP-Seq), microarrays, and other applications.
For specific notes and additional information, please see the product protocol PDF.
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