The EpiQuik™ Tissue Methylated DNA Immunoprecipitation (MeDIP) Kit is a complete set of essential components which uses a proprietary and unique procedure/composition to enrich methylated DNA from various mammalian tissues. In the assay, an antibody specific to methyl cytosine is used to immunoprecipitate methylated genomic DNA. The immunoprecipitated methylated fractions can be then used for a standard DNA detection. The kit is suitable for combining the specificity of methylated DNA immunoprecipitation with qualitative and quantitative PCR, and southern blot as well as DNA microarray.

WHY CHOOSE THE EPIQUIK™ TISSUE MEDIP KIT?

• Directly immunoprecipitate methylated fractions of DNA from tissue lysates.
• Highly efficient enrichment of methylated DNA: > 95%
• The fastest procedure available, which can be finished within 3 and a half hours.
• Strip microwell format makes the assay flexible: manual or high throughput.
• Columns for DNA purification are included: save time and reduce labor.
• Compatible with all DNA amplification-based approaches.
• Simple, reliable, and consistent assay conditions.

The EpiQuik™ Tissue Methylated DNA Immunopricipitation (MeDIP) Kit contains all reagents required for carrying out a successful methylated DNA immunoprecipitation directly from mammalian tissues. Particularly, this kit includes a ChIP-grade 5-methylcytosine antibody and a negative control normal mouse IgG. DNA in the cells is extracted, sheared, and added into the microwell immobilized with the antibody. DNA is released from the antibody-DNA complex, and purified through the specifically designed Fast-Spin Column. Eluted DNA can be used for various down-stream applications.

SCHEMATIC PROCEDURE:

CP1 (Wash Buffer)
CP2 (Antibody Buffer)
CP3 (Lysis Buffer)
CP4 (ChIP Dilution Buffer)
CP5 (DNA Release Buffer)
CP6 (Reverse Buffer)
CP7 (Binding Buffer)
CP8 (Elution Buffer)
Homogenizing Buffer
Normal Mouse IgG (1 mg/ml)*
Anti-5-Methylcytosine (1 mg/ml)*
Proteinase K (10 mg/ml)*
8-Well Assay Strips (with Frame)
8-Well Strip Caps
F-Spin Column
F-Collection Tube
User Guide

* For maximum recovery of the products, centrifuge the original vial after thawing prior to opening the cap.

1. If I am using kit P-2020, starting with purified DNA rather than cells, would I have to modify any steps in the protocol?
When using kit P-2020, purified DNA cannot be used as starting material. The washing buffer, lysis buffer, and reversing buffer in the kit are not suitable for purified DNA.

2. What are the differences between the EpiQuik Methylated DNA IP Kit (#P-2020) and the MethylAmp Methyl DNA Capture Kit (#P-1015)?
Kit P-2020 is specifically designed for immunoprecipitating mDNA fraction directly from tissues. The main advantage of P-2019/2020 is that no pre-isolation/purification of DNA is required if mammalian cells/tissues are used.

3. On page 10 of the user guide, does the "input" vial need to add 150 ul CP7 as the samples do?
Yes, CP7 should also be added into the "input" vial for DNA binding to the column.

4. Why would WGA amplify both DNA from sample and control IgG?
10% background from IgG precipitatation, after WGA, it will also be amplified. It is not necessary for microarray use. Input should be used.

5. How can I enrich 1-4µg of DNA, as required for microarray?
Amplify IPed DNA by WGA.

[User Guide]* *Always use the actual User Guide that shipped with your product. Is the above file locked? You can also request user guides by emailing info@epigentek.com along with your contact information and institution name.

[Material Safety Data Sheet]

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Roth, T. L. et. al. (May 1, 2009). Lasting epigenetic influence of early-life adversity on the BDNF gene. Biological Psychiatry. 65(9): 760-9. Full PDF Article