Human Control qPCR Primer Sets were designed to serve as positive or negative ChIP controls when performing chromatin immunoprecipitation with human samples. Each primer set is a mixture of forward and reverse primers that have been validated for qPCR of ChIP samples from multiple human cell lines.
In addition Chromatrap® supply forward and reverse primers on their own for common targets.
Chromatrap®’s Positive Primer Set 1 recognises the promoter region of the GAPDH gene that is associated with active transcription. A 69 base pair fragment is amplified with this primer set.
It is a positive gene target for the following: H3K4me1, H3K4me3, H3K27ac and RNA Pol II; among others.
Chromatrap®’s Positive Primer Set 2 amplifies a 134 base pair fragment of the imprinted H19 gene. It is a hypermethylated gene creating an appropriate positive target for methylation study.
It is a positive gene target for 5-methylcytosine (5mC).
Chromatrap®’s Positive Primer Set 3 recognises a region proximal to the promoter of the zinc finger DNA-binding protein, Myt1, that is present when transcription is repressed.
It is a positive gene target for the histone methylation mark H3K27me3 and EZH2.
Chromatrap®’s Positive Primer Set 4 amplifies a 72 base pair fragment of the protein-coding SAT2 gene.
This gene is present when transcription is repressed making it a positive gene target for the histone methylation mark H3K9me3.
Chromatrap®’s Positive Primer Set 5 recognises the promoter region of the protein-coding FkBP5 gene that has roles in glucocorticoid and progestin signalling pathways. It can regulate prostrate physiology in the presence of androgen (Magee et al. 2013). An 81 base pair fragment is amplified with this primer set.
It is a positive gene target for the male hormone receptor gene AR (Androgen Receptor) in the presence of testosterone.
Chromatrap®’s Positive Primer Set 6 recognises the promoter region of the DNA mismatch repair gene, MLH1. A 101 base pair product on human chromosome 3 is amplified by this primer set.
It is a positive gene target for the transcription factor Sin3a.
Chromatrap®’s Positive Primer Set 7 recognises the promoter region of the human B-Globin gene located on chromosome 11. A 121 base pair product is amplified with this primer set.
It is a positive gene target for the histone acetylation mark H3K14ac and RNA Pol II.
Chromatrap®’s Positive Primer Set 8 amplifies a 78 base pair fragment of the Interleukin-8 gene, found on human chromosome 4. IL-8 is a pro-inflammatory chemokine, whose increased expression correlates with the progression of cancer.
It is a positive gene target for the nuclear transcription factor NF-kB.
Chromatrap®’s Positive Primer Set 9 provides the necessary tools required for successful qPCR to validate your ChIP assay. It is a complete primer set including both forward and reverse primers, with sufficient primer mix supplied for a whole 96 well plate. Chromatrap®’s Positive Primer Set 9 is not suitable for use with non-human species.
Background:
Chromatrap®’s Positive Primer Set amplifies a 104 base pair fragment of the GREB1 (Growth regulation by estrogen in breast cancer 1) gene, found on human chromosome 2. GREB1 is an estrogen responsive gene that occurs in the estrogen-regulated pathway. It has roles in in hormone responsive tissues and in cancer.
It is a positive gene target for the histone modification H4K12ac in the presence of estrogen.
