Cells were crosslinked in 1% formadehyde for 15 min, with constant agitation, and quenched with 150mM glycine for 5 min. Micrococcal nuclease (New England Biolabs, Inc.) was used to fragment chromatin to a range of 200–400 bp. Chromatin was incubated with indicated antibody overnight at 4°C with constant agitation. Protein A Dynabeads was incubated with antibody-protein complexes for 2 hr at 4°C with constant agitation. After decrosslinking, DNA was isolated using a MinElute column (Qiagen). See Table S4 for primer sequence. For ChIP-qPCR, Sybr Green Taq 2× Master Mix (4309155, Life Technologies) was used to amplify DNA fragments.
Sybr green taq 2 master mix
Manufactured by Thermo Fisher Scientific
Sybr Green Taq 2x Master Mix is a pre-formulated solution containing Taq DNA polymerase, SYBR Green I dye, dNTPs, and reaction buffers optimized for real-time PCR applications. It is designed to simplify real-time PCR setup and improve consistency of results.
Automatically generated - may contain errors
Lab products found in correlation
2 protocols using sybr green taq 2 master mix
1
Chromatin Immunoprecipitation (ChIP) Protocol
2
Chromatin Immunoprecipitation (ChIP) Protocol
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