EpiTect ChIP qPCR Assay kit (Qiagen) was used for ChIP-qPCR assay. Briefly, the chromatin was isolated from fixed cells and was sonicated to an average size of 0.8–1.0 kb. The sample was then incubated with beads with the antibody for immunoprecipitation. The precipitated DNA was then subjected to qRT-PCR analysis. DNA Enrichment was calculated as a percentage of input DNA.
Epitect chip qpcr assay kit
Manufactured by Qiagen
Sourced in United States
The EpiTect ChIP qPCR Assay kit is a laboratory equipment product designed for chromatin immunoprecipitation (ChIP) and quantitative PCR (qPCR) analysis. The kit provides the necessary components to perform ChIP experiments and subsequent qPCR to detect and quantify specific DNA sequences bound by transcription factors or other DNA-binding proteins.
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4 protocols using epitect chip qpcr assay kit
1
ChIP-qPCR Assay Protocol Using EpiTect Kit
2
PTEN Promoter ChIP-qPCR Assay
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ChIP assay was performed using the EpiTect ChIP qPCR Assay Kit (Qiagen, Hilden, Germany) in accordance with the manufacturer's instruction manual. The bound PTEN promoter sequence was verified using RT-PCR (Real-Time polymerase chain reaction) assay with DNA fragments immunoprecipitated by anti-p300 antibodies (ab14984, Abcam, Cambridge, MA), anti-acetyl-histone H3 (06-599; Upstate Biotechnology, Lake Placid, NY, USA), or control rabbit IgG. The DNA fragments were amplified by quantitative RT-PCR using Direct SYBR Premix (Takara, Dalian, China) on the LightCycler 480 Real-Time PCR System (Roche Diagnostics). The RT-PCR primers are 5′-CGG GCG GTG ATG TGG C-3′ and 5′-GCC TCA CAG CGG CTC AAC TCT-3′.
3
ChIP-PCR Assay for SOX2, YAP, and WWTR1
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EpiTect ChIP qPCR Assay kit (#334211, Qiagen, Valencia, CA, USA) was used for ChIP-PCR assay. Brie y, the cells were xed for cross-linking, followed by isolation of chromatin. The chromatin DNA was subsequently fractionated to 0.8 to 1.0 kb by sonication, followed by incubation with SOX2 antibodybound beads. The DNA samples on the bead were then reverse cross-linked and collected. After puri cation and quanti cation, the Levels of YAP and WWTR1 promoter were determined by qRT-PCR.
4
ChIP-PCR Assay for SOX2, YAP, and WWTR1
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EpiTect ChIP qPCR Assay kit (#334211, Qiagen, Valencia, CA, USA) was used for ChIP-PCR assay. Brie y, the cells were xed for cross-linking, followed by isolation of chromatin. The chromatin DNA was subsequently fractionated to 0.8 to 1.0 kb by sonication, followed by incubation with SOX2 antibodybound beads. The DNA samples on the bead were then reverse cross-linked and collected. After puri cation and quanti cation, the Levels of YAP and WWTR1 promoter were determined by qRT-PCR.
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