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EZ-96 DNA Methylation MagPrep

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Cat #	Name	Size	Price	Quantity
D5040	EZ-96 DNA Methylation MagPrep	4 x 96 Rxns.	$742.80	- +
D5041	EZ-96 DNA Methylation MagPrep	8 x 96 Rxns.	$1,186.60	- +

Highlights

Proven procedure for complete bisulfite conversion of DNA.
High-throughput (96-well), automated desulphonation and recovery of bisulfite-treated DNA.
Recovered DNA is ideal for downstream analyses including PCR, endonuclease digestion, sequencing, microarrays, etc.

Documents

Supplemental Info

Product Description

The EZ-96 DNA Methylation MagPrep is a bisulfite conversion kit that features a high-throughput (96-well), simplified procedure that streamlines bisulfite conversion of DNA. The kit is based on the three-step reaction that takes place between cytosine and sodium bisulfite during which cytosine is converted into uracil. Desulphonation and clean-up of the converted DNA is performed while bound to the MagBinding Beads. The kit is designed to reduce template degradation and minimize DNA loss during treatment and clean-up, while ensuring complete conversion of the DNA. Purified, converted DNA is ideal for downstream analyses including PCR amplification, endonuclease digestion, sequencing, microarrays, etc.

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

Documents

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%
Equipment	Thermocycler with heated lid, heating element for 96-well plate, magnetic stand.
Input	500 pg - 2 µg of DNA
Processing Time	12-16 hours
Recovery	> 70%
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: 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.

Q2: 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.

Q3: 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.

Q4: What is the minimum DNA size that can be recovered?

> 50 bp.

Q5: 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	Price
C2003	Elution Plate	2 Plates	$21.50
C2002	Collection Plate	2 Plates	$24.90
C2005	Conversion Plate w/ Cover Foil	2 Plates/Foils	$10.30
D5040-3	M-Binding Buffer	250 ml	$129.80
D5040-5	M-Desulphonation Buffer	80 ml	$91.50
D5040-4	M-Wash Buffer	72 ml	$84.00
D5007-6	M-Elution Buffer	8 ml	$12.40
D5003-1	CT Conversion Reagent (96 Conversions)	1 Bottle	$77.90
D5041-6	M-Elution Buffer	40 ml	$37.10
D5002-2	M-Dilution Buffer	5.2 ml	$12.40
D4100-5-8	MagBinding Beads	8 mL	$101.20
D4100-5-16	MagBinding Beads	16 mL	$182.60