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The EpiQuik™ Chromatin Immunoprecipitation (ChIP) Kit is a convenient package of tools that allows the experimenter to perform chromatin immunoprecipitation (ChIP) at extraordinarily rapid speeds and consistency, superior to all other current ChIP methods available. The kit is ready-to-use and provides all the essential components needed to carry out a successful ChIP experiment. The EpiQuik™ ChIP kits are suitable for combining the specificity of immunoprecipitation with qualitative and quantitative PCR, MS-PCR, DNA sequencing, and southern blot as well as DNA microarray.

WHY CHOOSE THE EPIQUIK™ CHROMATIN IMMUNOPRECIPITATION CHIP KIT?

• The number one fastest procedure available. The entire procedure can be completed within an amazingly short 5 hour period with superb results.
• The ChIP procedure has been drastically simplified -- extremely short and easy to follow.
• Strip microwell format makes the assay flexible: manual or high throughput.
• Columns for DNA purification are included: save tremendous amounts of time and reduce unnecessary physical labor.
• Compatible with all DNA amplification-based approaches.
• Achieves very reliable and consistent assay conditions.

This kit includes a positive control antibody (RNA polymerase II), a negative control normal mouse IgG, and GAPDH primers that can be used as a positive control to demonstrate the efficacy of the kit reagents and protocol. RNA polymerase II is considered to be enriched in the GAPDH gene promoter that is expected to be undergoing transcription in most growing mammalian cells and can be immunoprecipitated by RNA polymerase II but not by normal mouse IgG.

In this ChIP, cells are cross-linked with formaldehyde and chromatin is extracted. The chromatin is then sheared and added into the microwell immobilized with affinity antibodies. Cross-linked DNA is released from antibody-captured protein-DNA complex, reversed and purified through the specifically designed Fast-Spin Column. Eluted DNA can be used for various down-stream applications.

SCHEMATIC PROCEDURE:	COMPARATIVE OVERVIEW:

CP1 (wash buffer)
CP2 (antibody buffer)
CP3A (lysis buffer)
CP3B (lysis buffer)
CP4 (ChIP dilution buffer)
CP5 (DNA release buffer)
CP6 (reverse buffer)
CP7 (binding buffer)
CP8 (elution buffer)
Protease inhibitor cocktail (100x)*
Normal mouse IgG (1 mg/ml)*
Anti-RNA polymerase II (1mg/ml)*
Proteinase K (10 mg/ml)*
Control primer (GAPDH)
Forward (20 µM)*
Reverse (20 µM)*
8-well assay strips (with frame)
8-well strip caps
F-spin column
F-collection tube
User guide

* Spin the solution down to the bottom before use.

1. Can IP be overnight?
Yes.

2. Is there any background DNA by mouse IgG IP?
Yes, a little < 2 ng.

3. Can any species Ab can be used for the IP?
Yes, if it is an IgG antibody.

4. Can kit P-2002 be used for frozen tissues?
Yes, but it is not as good as fresh samples.

5. How much volume of elution should be added into PCR reaction?
2 µl.

6. Are cell extracts containing 1%SDS compatible with CP4?
No, the SDS concentration in IP solution should be less than 0.1%.

7. Why is there no difference of CT values between the negative control and samples?
A. Increase washing time by one.
B. Check if antibody is chip grade.
C. There is no enrichment on the promoter.

8. Why is the difference between negative control and positive control not significant?
A. For conventional PCR, reduce PCR cycles to 35-36 or optimize the PCR cycles.
B. Increase washing time by one additional wash.

[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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