highlights
- Â Fastest 96-well bisulfite conversion kit for complete, high-throughput (96-well) bisulfite conversion of DNA for methylation analysis.
 Ready-to-use conversion reagent is added directly to DNA. -  High-yield, converted DNA is ideal for PCR, Methylation Specific PCR (MSP), arrays, library preps, Next-Generation sequencing, etc.
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The EZ-96 DNA Methylation-Lightning Kit is a rapid, high-throughput (96-well) bisulfite conversion kit for DNA methylation analysis. The streamlined workflow features ready-to-use Lightning Conversion Reagent and a combined DNA denaturation and bisulfite conversion process. Desulphonation and clean-up of the converted DNA is performed using a unique 96-well spin-plate. With the Deep-Well Kit (Cat. D5033), samples can be eluted in as little as 15 µl. High yield, converted DNA is ideal for PCR, array, and Next-Generation sequencing, etc.
| Applications | Purified, converted DNA is of high-quality and well-suited for downstream processes, including library preparation for Next-Generation sequencing, PCR amplification, etc. |
|---|---|
| Conversion | > 99.5% |
| Elution Volume | Deep-well: ≥ 15 µl Shallow-well: ≥ 30 µl |
| Equipment | Thermocycler with heated lid, swinging-bucket centrifuge with plate carriers. |
| Input | 100 pg – 2 µg of DNA. |
| Processing Time | 1.5 hours |
| Recovery | > 80% |
| Sample Source | Purified genomic DNA, endonuclease-digested DNA, linearized plasmid DNA, etc. DNA should be high-quality and RNA-free. |
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FAQs
ZymoTaq DNA Polymerase has been specifically designed for use in bisulfite amplification reactions. ZymoTaq is available as a stand-alone polymerase (E2001/E2002), PreMix (E2003/E2004), and qPCR PreMix (E2054/E2055).
Leaving the desulphonation buffer on the column longer than recommended will cause more degradation and subsequently result in lower yields.
Bisulfite conversion will work regardless of context, so the kits are compatible with genomic DNA derived from plants and other species with high non-CpG methylation levels.
Following bisulfite treatment of genomic DNA, nonmethylated cytosine residues are converted into uracil. The recovered DNA is typically A, U, and T-rich. The original base-pairing no longer exists. Instead, it is single-stranded with limited non-specific base-pairing at room temperature. The absorption coefficient at 260 nm resembles that of RNA. Use a value of 40 ÎĽg/ml for Ab260 = 1.0 when determining the concentration of the recovered bisulfite-treated DNA. To visualize, run converted DNA on agarose gel then chill the gel on ice for 30 minutes. The expected smear will be between 100-1500bp.
Poor conversion efficiency and low yields can be due to a variety of different experiment-specific conditions. Please contact Technical Support to discuss your specific experimental conditions and further troubleshoot with a product specialist.
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Kit Components
| Cat # | Name | Size |
|---|---|---|
| D5032-1 | Lightning Conversion Reagent | 15 ml |
| D5031-5 | L-Desulphonation Buffer | 40 ml |
| D5006-3 | M-Binding Buffer | 125 ml |
| D5007-6 | M-Elution Buffer | 8 ml |
| D5007-4 | M-Wash Buffer | 36 ml |
| C2001 | Silicon-A Plate | 2 Plates |
| C2004 | Zymo-Spin I-96 Plate | 2 Plates |
| C2005 | Conversion Plate w/ Cover Foil | 2 Plates/Foils |
| C2002 | Collection Plate | 2 Plates |
| C2003 | Elution Plate | 2 Plates |
EZ-96 DNA Methylation-Lightning Kit
| Cat # | Name | Size |
|---|---|---|
| D5032-E | EZ-96 DNA Methylation-Lightning Kit (Shallow-Well) (CE-IVD) | 2 x 96 Rxns. |
| D5033-E | EZ-96 DNA Methylation-Lightning Kit (Deep-Well) (CE-IVD) | 2 x 96 Rxns. |
| D5032 | EZ-96 DNA Methylation-Lightning Kit (Shallow-Well) | 2 x 96 Rxns. |
| D5033 | EZ-96 DNA Methylation-Lightning Kit (Deep-Well) | 2 x 96 Rxns. |