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SimpleChIP ® Universal qPCR Master Mix
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The SimpleChIP® Universal qPCR Master Mix is an optimized 2X reaction mix for real-time qPCR detection and quantitation of target DNA sequences using the SYBR/FAM channel of most real-time qPCR instruments. It contains Hot Start Taq DNA Polymerase and has been formulated with a unique passive reference dye that is compatible across a variety of instrument platforms (including those that require a high or low ROX reference signal). It also features dUTP for carryover prevention and a non-fluorescent, visible dye to monitor reaction setup. This dye does not spectrally overlap with fluorescent dyes used for qPCR and will not interfere with real-time detection.
This product is provided in 1 ml volumes sufficient for preparation of 100 qPCR reactions, and is compatible with both enzymatic and sonication-fragmented DNA samples from SimpleChIP® enzymatic and sonication ChIP kits. This master mix formulation is supplied at 2X concentration and contains all PCR components required for amplification and quantitation of DNA, except primers and a DNA template.
Specificity/Sensitivity
Species Reactivity:
All Species Expected
This product is stable for 12 months when stored at -20°C and protected from light. It is stable for up to 30 freeze/thaw cycles and can be stored at 4°C, protected from light, for up to 1 month.
参考图片
Figure 3. CUT&RUN and ChIP assays were performed with HCT 116 cells and CTCF (D31H2) XP® Rabbit mAb #3418 or Rabbit (DA1E) mAb IgG XP® Isotype Control (CUT&RUN) #66362, using CUT&RUN Assay Kit #86652 (left panel) or the SimpleChIP® Plus Enzymatic Chromatin IP Kit (Magnetic Beads) #9005 (right panel). The enriched DNA was quantified by real-time PCR using this SimpleChIP® Universal qPCR Master Mix, human c-Myc promoter primers, SimpleChIP® Human H19/Igf2 ICR Primers #5172, and SimpleChIP® Human α Satellite Repeat Primers #4486. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the total amount of input chromatin, which is equivalent to one.
FIGURE 1. Amplification plot (left) and regression curve (right) of a serial dilution of input DNA samples. Input DNA was prepared and purified from HCT 116 cells using SimpleChIP® Plus Enzymatic Chromatin IP Kit (Magnetic Beads) #9005. Quantitative PCR was performed with 20 (red line), 4 (orange line), 0.8 (purple line), and 0.016 ng (blue line) of input DNA and SimpleChIP® Human RPL30 Exon 3 Primers #7014. Data is shown for two replicates.
FIGURE 2. Corresponding single melt peak of primers. Quantitative PCR was performed with input DNA sample from HCT 116 cells prepared by SimpleChIP® Plus Enzymatic Chromatin IP Kit (Magnetic Beads) #9005 and either SimpleChIP® Human α Satellite Repeat Primers #4486 (red line), SimpleChIP® Human RPL30 Exon 3 Primers #7014 (orange line), SimpleChIP® Human CaMK2D Intron 3 Primers #5111 (purple line), or human c-MYC promoter primers (blue line). Data for two replicates is shown for each primer set.