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EZ-96 DNA Methylation-Lightning MagPrep
D5046 / D5047 / D5046-E / D5047-E
EZ-96 DNA Methylation-Lightning MagPrep
- 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.
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 firstname.lastname@example.org
|Applications||Purified, converted DNA is of high-quality and well-suited for downstream processes, including library preparation for Next-Generation sequencing, PCR amplification, etc.|
|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|
|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 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.
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.
“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)
|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|