EZ DNA Methylation Kit

EZ DNA Methylation Kit

Cat#NameSize
D5001EZ DNA Methylation Kit50 Rxns.
D5002EZ DNA Methylation Kit200 Rxns.

Documents

Protocol:

SDS (MSDS):

App Note:

Description

Highlights

  • Streamlined, proven procedure for bisulfite conversion of DNA.
  • Desulphonation and recovery of bisulfite-treated DNA with a spin column.
  • Recovered DNA is ideal for downstream analyses including PCR, endonuclease digestion, sequencing, microarrays, etc.

Description

The EZ DNA Methylation Kit features a simplified procedure that streamlines bisulfite treatment of DNA. This kit is the original bisulfite conversion kit from Zymo Research. The EZ DNA Methylation Kit is based on the three-step reaction that takes place between cytosine and sodium bisulfite during which cytosine is converted into uracil. Innovative desulphonation technologies eliminate otherwise cumbersome precipitations. 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 library preparation for Next-generation sequencing, PCR amplification, endonuclease digestion, sequencing, microarrays, etc. The EZ DNA Methylation Kit is recommended for use with Illumina Infinium MethylationEPIC BeadChip array.

Applicable For

Purified, converted DNA is of high-quality and well-suited for downstream processes, including library preparation for Next-generation sequencing, PCR amplification, etc. This kit is recommended for use with Illumina Infinium MethylationEPIC BeadChip array.

Conversion

>99%

Elution Volume

≥10 µl

Equipment

Microcentrifuge and thermocycler with heated lid

Input

500 pg – 2 µg of DNA

Processing Time

12-16 hours

Recovery

>80%

Sample Source

Purified genomic DNA, endonuclease-digested DNA, linearized plasmid DNA, etc. DNA should be high-quality and RNA-free.

Q1: 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.

Q2: Tips for bisulfite primer design?

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

Q3: 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).

Q4: 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.

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.

99 / 100
Bioz Stars

The integrated genomic landscape of thymic epithelial tumors

Cancer Cell – Published 12 Feb 2019

The Illumina Infinium HM450 array ( ) was used to assay 117 TCGA TET samples using standard protocols.. Briefly, genomic DNA (1000 ng) for each sample was treated with sodium bisulfite, recovered using the Zymo EZ DNA methylation kit (Zymo Research, Irvine, CA) according to the manufacturer’s specifications and eluted in 18 ul volume.. After passing quality control, bisulfite-converted DNA samples were whole genome amplified followed by enzymatic fragmentation and hybridized overnight

DNA methylation patterns separate senescence from transformation potenti…

Cancer Cell – Published 12 Feb 2019

Genomic DNA quality was assessed by low concentration agarose gel (0.6%) electrophoresis and spectrometry of OD260/280 and OD 260/230 ratio.. DNA bisulfite conversion was carried out using EZ DNA Methylation Kit (Zymo Research) by following manufacturer’s manual with modifications for Illumina Infinium Methylation Assay.. Briefly, 0.5 – 1.0 ug of genomic DNA was first mixed with 5 ul of M-Dilution Buffer and incubate at 37°C for 15 minutes and then mixed with 100 ul

TETs Regulate Proepicardial Cell Migration through Extracellular Matrix …

Cell Rep – Published 07 Feb 2019

Peak calling and analysis of read density in peak regions were performed by macs14 1.4.2 with default parameters ( ).. Bisulfite sequencing was performed using the EZ DNA Methylation-Direct kit (Zymo Research).. Embryonic hearts at 48 hpf were dissected (n = 4 per condition).

Cat # Name Size
D5001-1 CT Conversion Reagent 1 tube for 10 Conversions
D5001-2 M-Dilution Buffer 1.3 ml
D5002-2 M-Dilution Buffer 5.2 ml
D5001-3 M-Binding Buffer 20 ml
D5002-3 M-Binding Buffer 80 ml
D5001-4 M-Wash Buffer 6 ml
D5002-4 M-Wash Buffer 24 ml
D5001-5 M-Desulphonation Buffer 10 ml
D5002-5 M-Desulphonation Buffer 40 ml
D5001-6 M-Elution Buffer 1 ml
D5002-6 M-Elution Buffer 4 ml
C1001-50 Collection Tubes 50 Pack
C1004-50 Zymo-Spin IC Columns 50 Pack
  • Catalog#: D5001 /D5002
  • Package Length (in Inches): 7 /9
  • Package Width (in Inches): 5 /6.5
  • Package Height (in Inches): 4 .5/6
  • Package Weight (in Pounds): 0.8 /2.3
  • Size: 50 Rxns /200 Rxns
  • Unit Standard: Metric
  • Volume Units: Milliliters