EZ-96 DNA Methylation-Lightning MagPrep

D5046 / D5047


EZ-96 DNA Methylation-Lightning MagPrep

D5046 / D5047

Cat # Name Size Price Quantity
D5046 EZ-96 DNA Methylation-Lightning MagPrep 4 x 96 Rxns. $696.00
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D5047 EZ-96 DNA Methylation-Lightning MagPrep 8 x 96 Rxns. $1,126.00
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Documents


Discover the fastest bisulfite conversion kit with the most efficient method for complete bisulfite conversion of DNA for methylation analysis.

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.
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.


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.

Q1: Tips for bisulfite primer design?
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 / 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.




“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.”

- Marco M. (UCLA)


Cat # Name Size Price
D4100-2-16 MagBinding Beads 16 ml $166.00
D4100-2-8 MagBinding Beads 8 ml $92.00
D5040-3 M-Binding Buffer 250 ml $105.00
D5040-4 M-Wash Buffer 72 ml $68.00
D5007-6 M-Elution Buffer 8 ml $10.00
D5041-6 M-Elution Buffer 40 ml $30.00
D5046-5 L-Desulphonation Buffer 80 ml $74.00
D5032-1 Lightning Conversion Reagent 15 ml $136.00
C2002 Collection Plate 2 Plates $22.00
C2003 Elution Plate 2 Plates $19.00
C2005 Conversion Plate w/ Cover Foil 2 Plates/Foils $10.00