SimpleChIP® Human HES5 Upstream Primers #59885
- ChIP
Supporting Data
| REACTIVITY | H |
Application Key:
- ChIP-Chromatin Immunoprecipitation
Species Cross-Reactivity Key:
- H-Human
Product Information
Product Description
Product Usage Information
Directions for Use:
1. Label the appropriate number of PCR tubes or PCR plates
compatible with the model of real-time PCR machine to be used.
PCR reactions should be performed in duplicate and should
include a tube with no DNA to control for contamination, and
a serial dilution of a 2% total input chromatin DNA (undiluted,
1:5, 1:25, 1:125), which is used to create a standard curve and
determine amplification efficiency.
2. Add 2 μl of the appropriate ChIP DNA sample to each tube or
well of the PCR plate.
3. Prepare a master PCR reaction mix as described below. Add
enough reagents for two extra reactions to account for loss of
volume. Add 18 μl of the master PCR reaction mix to each PCR
reaction tube or well of the PCR plate.
Reagent Volume for 1 PCR Reaction (20 μl)
Nuclease-free H2O 6 μl
5 μM SimpleChIP® Primers 2 μl
SimpleChIP® Universal qPCR Master Mix #88989 10 μl
4. Start the following PCR reaction program:
a. Initial Denaturation: 95°C for 3 min
b. Denaturation: 95°C for 15 sec
c. Anneal and Extension: Primer-specific temp. for 60 sec
d. Repeat steps b and c for a total of 40 cycles.
5. Analyze quantitative PCR results using software provided with the real-time PCR machine.
Storage
Protocol
Specificity / Sensitivity
Species Reactivity:
Background
- Orlando, V. (2000) Trends Biochem Sci 25, 99-104.
- Kuo, M.H. and Allis, C.D. (1999) Methods 19, 425-33.
- Agalioti, T. et al. (2000) Cell 103, 667-78.
- Soutoglou, E. and Talianidis, I. (2002) Science 295, 1901-4.
- Mikkelsen, T.S. et al. (2007) Nature 448, 553-60.
- Lee, T.I. et al. (2006) Cell 125, 301-13.
- Weinmann, A.S. and Farnham, P.J. (2002) Methods 26, 37-47.
- Wells, J. and Farnham, P.J. (2002) Methods 26, 48-56.
Limited Uses
Except as otherwise expressly agreed in a writing signed by a legally authorized representative of CST, the following terms apply to Products provided by CST, its affiliates or its distributors. Any Customer's terms and conditions that are in addition to, or different from, those contained herein, unless separately accepted in writing by a legally authorized representative of CST, are rejected and are of no force or effect.
Products are labeled with For Research Use Only or a similar labeling statement and have not been approved, cleared, or licensed by the FDA or other regulatory foreign or domestic entity, for any purpose. Customer shall not use any Product for any diagnostic or therapeutic purpose, or otherwise in any manner that conflicts with its labeling statement. Products sold or licensed by CST are provided for Customer as the end-user and solely for research and development uses. Any use of Product for diagnostic, prophylactic or therapeutic purposes, or any purchase of Product for resale (alone or as a component) or other commercial purpose, requires a separate license from CST. Customer shall (a) not sell, license, loan, donate or otherwise transfer or make available any Product to any third party, whether alone or in combination with other materials, or use the Products to manufacture any commercial products, (b) not copy, modify, reverse engineer, decompile, disassemble or otherwise attempt to discover the underlying structure or technology of the Products, or use the Products for the purpose of developing any products or services that would compete with CST products or services, (c) not alter or remove from the Products any trademarks, trade names, logos, patent or copyright notices or markings, (d) use the Products solely in accordance with CST Product Terms of Sale and any applicable documentation, and (e) comply with any license, terms of service or similar agreement with respect to any third party products or services used by Customer in connection with the Products.