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EZ DNA Methylation-Lightning Kit Sample

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Highlights

Fastest bisulfite conversion kit for 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.

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Supplemental Info

Product Description

The EZ DNA Methylation-Lightning Kit can rapidly bisulfite convert and purify DNA in less than 1.5 hours. A ready-to-use conversion reagent streamlines this process – simply add the reagent directly to the sample and incubate. Desulphonation and clean-up of the converted DNA is performed on a spin-column, allowing an elution volume as low as 10 µl. The kit achieves high conversion efficiency of unmodified cytosines into uracil, ensuring accurate downstream methylation analysis.

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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	≥ 10 µl
Equipment	Thermocycler with heated lid and microcentrifuge.
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: 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).

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: Tips for bisulfite primer design?

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

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

> 50 bp.

Q6: How to quantify / visualize converted DNA?

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 μg/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.

Q7: 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.

Q8: How long is bisulfite converted DNA stable at -20 °C?

Converted DNA should be eluted in M-Elution Buffer to keep the converted DNA stable for long term storage. If stored properly for long term (<-20C), the samples should last longer than a month. Minimize freeze/thawing to keep the bisulfite converted DNA stable.

Cat #	Name	Size	Price
C1004-50	Zymo-Spin IC Columns	50 Pack	$61.80
C1001-50	Collection Tubes	50 Pack	$17.50
D5002-4	M-Wash Buffer	24 ml	$31.80
D5001-4	M-Wash Buffer	6 ml	$12.80
D5005-3	M-Binding Buffer	30 ml	$25.40
D5002-6	M-Elution Buffer	4 ml	$12.80
D5001-6	M-Elution Buffer	1 ml	$12.80
D5006-3	M-Binding Buffer	125 ml	$73.90
D5030-5	L-Desulphonation Buffer	10 ml	$19.10
D5031-5	L-Desulphonation Buffer	40 ml	$53.50
D5030-1	Lightning Conversion Reagent	1.5 ml	$29.30

“Results were as good as with the formerly used EZ DNA Methylation Gold Kit but significantly reduced total time for analyses. More flexibility due to the possibility of up to 20h storage of the samples after bisulfite conversion before the cleaning procedure especially for part-time employees.”

- Sabrina S. (UK-Aachen)

“The conversion reagent no longer has to be made up from individual components and dissolved - it is supplied ready-made. The conversion reagent can be stored at room temperature - there is no need to use up all 10 uses in one month. This makes it more flexible, and avoids wastage. Shorter protocol saves time.”