About EZ-96 DNA Methylation-Lightning™ Kit
The EZ-96 DNA Methylation-Lightning™ Kit features high-throughput (96-well) bisulfite treatment and conversion of DNA for methylation analysis. Key to the fast workflow is the ready-to-use Lightning Conversion Reagent. No preparation is necessary, simply add this unique reagent to a DNA sample, wait about an hour, and let the reaction proceed to completion. DNA denaturation and bisulfite conversion processes are combined with added heat to facilitate rapid denaturation. Desulphonation and clean-up of the converted DNA is performed using a unique 96-well spin-plate. High yield, converted DNA is ideal for PCR, array, bisulfite and next generation sequencing, etc.
|Learn More About Bisulfite Conversion||Click to see which kit is right for you|
|DNA Recovery||> 80%|
|Conversion Efficiency||>99.5% of non-methylated cytosine residues are converted to uracil; >99.5% protection of methylated cytosines.|
|Optimal DNA Input||Samples containing between 100 pg to 2 µg of DNA. For optimal results, the amount of input DNA should be from 200 to 500 ng.|
Add 130 µl of Lightning Conversion Reagent to 20 µl of a DNA sample in a Conversion Plate. Mix the samples by pipetting up and down.
Note: If the volume of DNA is less than 20 µl, compensate with water.
Seal the plate with the provided film. Transfer the Conversion Plate to a thermal cycler and perform the following steps:
1. 98°C for 8 minutes
2. 54°C for 60 minutes
3. 4°C storage for up to 20 hours
Note: The 4°C storage step is optional.
Add 400 µl of M-Binding Buffer to the wells of a Silicon-A™ Binding Plate mounted on a Collection Plate.
Transfer the samples from the Conversion Plate (Step 2) to the wells of the Silicon-A™ Binding Plate. Mix by pipetting up and down.
Centrifuge at ≥ 3,000 x g (5,000 x g max.) for 5 minutes. Discard the flow-through.
Add 400 µl of M-Wash Buffer to each well of the plate. Centrifuge at ≥ 3,000 x g for 5 minutes.
Add 200 µl of M-Desulphonation Buffer to each well and allow the plate to stand at room temperature (20-30°C) for 15-20 minutes. After the incubation, centrifuge at ≥ 3,000 x g for 5 minutes. Discard the flow-through.
Add 400 µl of M-Wash Buffer to each well of the plate. Centrifuge at ≥ 3,000 x g for 5 minutes. Discard the flow-through. Add another 400 µl of M-Wash Buffer and centrifuge for 10 minutes.
Place the Silicon-A™ Binding Plate onto an Elution Plate. Add 30 µl of M-Elution Buffer directly to each well. Wait 5 minutes, then centrifuge at ≥ 3,000 x g for 3 minutes to elute the DNA.
The DNA is ready for immediate analysis or can be stored at or below -20°C for later use. For long term storage, store at or below -70°C. We recommend using 1-4 µl of eluted DNA for each PCR, however, up to 30 µl can be used if necessary. The elution volume can be < 30 µl depending on the requirements of your experiments, but small elution volumes will yield higher DNA concentrations.