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BIOPANEL® methylation detection kit for human pluripotent stem cells (Light Cycler™ 480 RT PCR)

Rapid, high-throughput assay for quantifying methylation to determine pluripotency state of cells using Light Cycler™ 480 RT PCR
 
ENZ-45007-0001 96 wells 552.00 USD
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  • High-throughput quantification of DNA methylation in RAB25, NANOG, PTPN6, MGMT, GBP3, and LYST gene regions in human stem cells or for indicating pluripotency in stem cell lines
  • Convenient 96-well assay for bisulfite-free methylation percent quantitation of specific DNA regions
  • Easy to follow instructions
The BIOPANEL® methylation detection kit for pluripotent human stem cells provides a simple and bisulfite-free procedure for rapid, DNA methylation assessment of: RAB25, NANOG, PTPN6, MGMT, GBP3, and LYST in any cell type. The reagents in the plate (compatible with Light Cycler™ 480 RT PCR) are already premixed and optimized for robust amplification and detection. Simply add DNA into the appropriate well and then quantitate via real-time PCR.
ENZ-45007 Fig1
Figure demonstrates the work flow of non-methylated DNA (open circles) and methylated (closed circles). In both scenarios an Experimental Reaction and Reference Reaction is required. The Experimental Reaction is MSRE digested while the Reference Reactions is not. DNA from both samples are then used for real-time PCR. Following real-time PCR, amplification plots (bottom panel) demonstrate that non-methylated DNA exhibits a large Ct value difference for Experimental Reaction and Reference Reaction. This type of data is used for the basis for methylation determination.
ENZ-45007 Fig4
Figure shows an example of difference in methylation percentages for various genes including, RAB25, NANOG, PTPN6, MGMT, GBP3 and LYST. Human differentiated DNA (yellow stripped bars) and human stem cell DNA (white bars) percentages were calculated following Ct calculations. This information helps identify if a cell is in a pluripotent state.
ENZ-45007 Fig2
Experimental set-up for duplicate sampling
ENZ-45007 Fig3
Experimental set-up for triplicate sampling
Please mouse over
ENZ-45007 Fig1 ENZ-45007 Fig4 ENZ-45007 Fig2 ENZ-45007 Fig3

Product Specification

Use/Stability:With proper storage, good for 6 months upon receipt.
 
Shipping:Shipped on Dry Ice
 
Long Term Storage:-20°C
 
Kit/Set Contains:BIOPANEL® Plate for Light Cycler™
 
Scientific Background:Pluripotency refers to the potential of a stem cell to differentiate into any of the three germ layers; endoderm, mesoderm or ectoderm. Pluripotent cells have epigenetic signatures that reflect their ability to generate multiple cell types. Different methylation patterns in gene regions vary between pluripotent and differentiated cells as a result of processes such as development, carcinogenesis, genomic imprinting disorders, and cell reprogramming. In human pluripotent cells, gene promoter regions in the RAB25, NANOG, and PTPN6 genes have been shown to maintain low levels of DNA methylation compared to differentiated cell types. Conversely, gene promoter regions of MGMT, GBP3, and LYST have been shown to maintain high levels of methylation in pluripotent cells compared to differentiated cell types.
 

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