DNA Bisulfite Conversion Bisulfite conversion is a technique in which cytosine is converted to uracil using sodium bisulfite, while 5-methylcytosine (5-mC) remains intact. Bisulfite and sodium bisulfite-based methods are common approaches used to study DNA methylation and help in preparing genomic DNA for gene-specific DNA methylation analysis. Bisulfite conversion is often followed by downstream applications such as PCR or sequencing. Unmethylated cytosine residues are deaminated to uracil and methylated cytosine (5-mC) residues remain unaffected, enabling PCR amplification to recognize uracils as thymines and 5-mC or 5-hmC as cytosines. This allows researchers to differentiate between methylated and unmethylated cytosine residues, offering single-nucleotide resolution information about the methylated areas of DNA. Complete conversion is necessary for a successful DNA methylation study, along with reducing the amount of DNA degradation that will occur as a result of the typically harsh chemical reaction. Popular downstream methods for analyzing DNA methylation on a gene-specific basis following bisulfite treatment include bisulfite sequencing, methylation specific PCR (MS-PCR), and methylation-based microarrays. Considering your downstream application can help in determining which bisulfite conversion method is ideal, especially due to the variation in post-conversion fragment size. The BisulFlash DNA Modification Kit is geared specifically toward real-time MSP whereas the best-selling Methylamp DNA Modification Kit is designed to be compatible with Illumina workflows and next-generation sequencing (NGS). If a quicker and easier bisulfite conversion alternative is desired prior to sequencing, the BisulFlash DNA Bisulfite Conversion Easy Kit can be completed in just 60 minutes and uses a liquid bisulfite conversion reagent for convenience.Beginning with high quality DNA is critical, especially because the acidic nature of the conversion process will fragment the DNA. For samples containing low amounts of DNA, the Methylamp Whole Cell Bisulfite Modification Kit is recommended, as it can directly convert from cells, tissues, blood, and other starting materials. When handling large scale bisulfite conversion experiments, high-throughput options are critical in order to save time and reduce cost. In this case, the BisulFlash DNA Bisulfite Conversion Mag-96 Kit is specifically streamlined for high-throughput and uses a magnetic bead-based 96-well format. For a detailed comparison of bisulfite conversion kits, see the table below.Codename\"Classic\"\"Fast\"\"Direct\"PurposeChemical-based DNA bisulfite conversionFast heat-based DNA bisulfite conversionDirectly convert from cells, tissues, saliva, urine, or bloodKey PointsIllumina workflow compatibleMost robust and reliableIdeal for bisulfite sequencing, NGS, and various MSPExtremely fastIdeal for real-time MSPOptional sequencing protocolEliminates separate DNA isolation stepIdeal for samples containing low quantities of DNAStarting MaterialDNADNAcells, tissue, bloodLowest Input Amount50 pg200 pg100 cells, 1µl bloodTotal Protocol Time 2 hours30 minutes 3 hoursElution Volume8-20µl10-20µl8-20µlConversion Efficiency99.9%99.9%99.9%Desulfonation/Clean-UpRecovery 75% 75% 85%Post-Conversion DNA Fragment Size200-2000 bp; Avg. Peak of 800 bp100-400 bp; Avg. Peak of 250 bp200-2000 bp; Avg. Peak of 800 bpVessel FormatSpin ColumnSpin ColumnSpin Column Lead TimeExpress Only (2 Days or Less)Research KitsInput Type DNA(8)Research Area DNA Methylation(8) Next Gen Sequencing(1)Target Application Sample Modification(8) Library Construction(1) Sample Isolation(0)Vessel Format Columns/Tubes(7) 96-Well Plate(0)

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