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EZ-96
DNA Methylation-Gold Kit™ (Shallow Well Format)
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Complete, high-throughput (96-well) bisulfite conversion
of GC-rich DNA in less than 3 hours. |
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A coupled DNA denaturation/bisulfite treatment step streamlines
the conversion of unmethylated cytosines into uracil. |
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DNA
precipitations are omitted. Instead, DNA is cleaned and
desulphonated in a single step. |
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Eluted,
ultra-pure DNA is ideal for use in subsequent molecular-based
analyses.
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SPECIFICATIONS
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Format: Rapid heat denaturation of DNA followed
by sodium bisulfite modification, Spin Column. |
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Recovery Volume: Purified,
bisulfite treated DNA is recovered in ≥ 10 µl provided
elution buffer. |
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Input
DNA: 500 pg - 2 µg DNA per treatment
with 200 - 500 ng being optimal. |
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Sample
Sources: Plasmid, genomic, and endonuclease
digested DNAs. |
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Equipment:
Microcentrifuge and thermalcycler, centrifuge w/ microplate
carriers (EZ-96 DNA Methylation-Gold Kit™) |
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DESCRIPTION
The 96-well format EZ-96 DNA Methylation-Gold
Kit™ is a refinement of our popular EZ-96
DNA Methylation Kit™. The EZ-96 DNA Methylation-Gold
Kit™ integrates the DNA denaturation and bisulfite
conversion processes into one-step. This is accomplished
using temperature denaturation to replace chemical denaturation
with sodium hydroxide in the previous protocol. Also, the
kit has been streamlined for high yield recovery of DNA
following DNA bisulfite conversion. Both kits are based
on a three-step reaction process between cytosine and sodium
bisulfite resulting in cytosine being converted into uracil.
The EZ-96 DNA Methylation-Gold™ and EZ-96 DNA Methylation™
Kits share innovative in-column desulphonation technology
that eliminates cumbersome DNA precipitation steps while
providing researchers consistent results every time. The
kits have been designed to minimize template degradation,
loss of DNA during treatment and clean-up, and to provide
complete conversion of unmethylated cytosines. Recovered
DNA is ideal for PCR amplification for downstream analyses
including restriction endonuclease digestion, sequencing,
microarrays, etc.
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DNA
sequencing results after bisulfite treatment. DNA
with methylated CmpG (at nucleotide position #5) was
processed using the EZ DNA Methylation-Gold
Kit™. The recovered DNA was amplified
by PCR and then sequenced directly. The methylated
cytosine at position #5 remained intact while the
unmethylated cytosines (i.e., positions #7, 9, 11,
14 and 15) were completely converted into uracil following
bisulfite treatment and were detected as thymine following
PCR. |
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ORDERING
INFORMATION:
| Components
Available Separately |
| CT
Conversion Reagent |
D5001-1 |
10
Conv. |
$6.10 |
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| D5003-1 |
96
Conv. |
$55.00 |
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| M-Dilution
Buffer-Gold |
D5005-2 |
1.5
ml |
$3.00 |
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| D5006-2 |
7
ml |
$6.50 |
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| M-Binding
Buffer-Gold |
D5005-3 |
30
ml |
$17.00 |
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| D5006-3 |
125
ml |
$50.00 |
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| M-Wash
Buffer |
D5001-4 |
6
ml |
$7.00 |
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| D5002-4 |
24
ml |
$21.00 |
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| D5007-4 |
36
ml |
$32.00 |
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| M-Desulphonation
Buffer |
D5001-5 |
10
ml |
$12.00 |
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| D5002-5 |
40
ml |
$36.00 |
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| M-Elution
Buffer |
D5001-6 |
1
ml |
$1.50 |
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| D5002-6 |
4
ml |
$4.50 |
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| M-Dissolving
Buffer-Gold |
D5005-6 |
500
µl |
$5.00 |
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| D5006-6 |
1.2
ml |
$12.00 |
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| Zymo-Spin
IC Columns
(capped) |
C1004-50
|
50
columns |
$46.00 |
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| C1004-250
|
250
columns |
$197.00 |
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| Collection
Tubes |
C1001-50 |
50
tubes |
$5.00 |
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| C1001-500 |
500
tubes |
$30.00 |
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| C1001-1000 |
1,000
tubes |
$50.00 |
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| Zymo-Spin
I-96 Filtration Plates |
C2004 |
2
plates |
$124.00 |
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Silicon-A Plates |
C2001 |
2
plates |
$112.00 |
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| Conversion
Plates w/Piercable
Cover Film |
C2005 |
2
plates |
$5.00 |
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| Collection
Plates |
C2002 |
2
plates |
$18.00 |
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| Elution
Plates |
C2003 |
2
plates |
$15.00 |
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RELATED PRODUCTS:
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Download
Data Sheet |
