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EZ-96 DNA Methylation-Lightning MagPrep

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Cat #	Name	Size
D5046	EZ-96 DNA Methylation-Lightning MagPrep	4 x 96 Rxns.
D5047	EZ-96 DNA Methylation-Lightning MagPrep	8 x 96 Rxns.
D5046-E	EZ-96 DNA Methylation Lightning MagPrep	4 x 96 Rxns.
D5047-E	EZ-96 DNA Methylation Lightning MagPrep	8 x 96 Rxns.

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Highlights

High-throughput procedure for automated rapid and complete 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.

Documents

Supplemental Info

Product Description

The EZ-96 DNA Methylation-Lightning MagPrep Kit is the fastest magnetic bead-based DNA bisulfite conversion kit for high-throughput and automated methylation analysis. No preparation is necessary with the ready-to-use conversion reagent - simply add this unique reagent to a DNA sample and incubate the reaction for about one hour. Desulphonation and clean-up of the converted DNA is performed while bound to the MagBinding Beads. This magnetic bead-based bisulfite conversion kit produces the highest yields of bisulfite-converted DNA that is ideal for PCR, arrays, library preps, and Next-Generation sequencing.

For automation scripts and support, email automation@zymoresearch.com

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Technical Specifications

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	≥ 25 µl
Equipment	Thermocycler with heated lid, heating element for 96-well plate, magnetic stand.
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.
Supplemental Info	Bisulfite Beginner Guide Controls for DNA Methylation Assays Bisulfite Conversion for Illumina Methylation Assays Bisulfite Primer Design Tool Link

Resources

Q1: Tips for bisulfite primer design?

https://www.zymoresearch.com/pages/bisulfite-beginner-guide
https://www.zymoresearch.com/pages/bisulfite-primer-seeker

Q2: Is an incubation with desulphonation buffer for longer than 20 minutes recommended?

Leaving the desulphonation buffer on the column longer than recommended will cause more degradation and subsequently result in lower yields.

Q3: Does bisulfite conversion only occur in a CpG context?

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.

Q4: Which polymerase is recommended for amplification from bisulfite converted DNA?

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).

Q5: What is the minimum DNA size that can be recovered?

> 50 bp.

Q6: How to quantify converted DNA?

For best results, keep the method of quantification consistent before and after bisulfite treatment:

If quantifying with a NanoDrop, use dsDNA settings (50 μg/ml for Ab260 = 1.0) before treatment and use RNA settings (40 μg/ml for Ab260 = 1.0) after treatment.
If quantifying with Qubit, use a dsDNA assay before treatment and use a ssDNA assay after treatment.

Following bisulfite treatment of 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, the converted DNA is single stranded with limited non-specific base-pairing at room temperature. Therefore, traditional dsDNA methods of quantification will not accurately represent the amount of recovered bisulfite converted DNA.

Q7: How to visualize converted DNA?

Following bisulfite treatment, DNA will be single stranded with limited non-specific base pairing at room temperature. To visualize, run the converted DNA on an agarose gel then chill the gel on ice or in an ice bath for 30 minutes. This will force enough base-pairing to allow intercalation of the ethidium bromide for the DNA to be visible. If using a Bioanalyzer or TapeStation instrument, use RNA kits and reagents to visualize the converted DNA.

Q8: What leads to poor conversion efficiency/ low yields?

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.

Q9: How long is bisulfite converted DNA stable at -20 °C?

Converted DNA eluted in M-Elution Buffer can generally be stored at -20°C for 1-3 months. If longer term storage is necessary, we recommend storing at or below -70°C if possible. Bisulfite converted DNA is less stable than dsDNA; for best results, minimize freeze-thawing of converted DNA and use as soon as possible for downstream analysis.

Cat #	Name	Size
D4100-5-8	MagBinding Beads	8 mL
D4100-5-16	MagBinding Beads	16 mL
D5041-6	M-Elution Buffer	40 ml
D5007-6	M-Elution Buffer	8 ml
D5040-4	M-Wash Buffer	72 ml
D5040-3	M-Binding Buffer	250 ml
D5032-1	Lightning Conversion Reagent	15 ml
D5046-5	L-Desulphonation Buffer	80 ml
C2005	Conversion Plate w/ Cover Foil	2 Plates/Foils
C2002	Collection Plate	2 Plates
C2003	Elution Plate	2 Plates

“It combines the speed of the lightning kit with the use of magnetic beads. This allows the full automation of bisulfite treatment. This is what we were looking for.”