This product is no longer available in the United States. For a similar product, please see BisulFlash DNA Modification Kit





The Methylamp™ DNA Modification Kit is a complete set of essential components which enables the experimenter to perform DNA methylation analysis using Epigentek's uniquely simplified and streamlined bisulfite method. The entire procedure can be completed within a mere 1 hour and 55 minutes and produces far superior results than any competitor kits. The Methylamp™ DNA Modification Kit is suitable for MS-PCR, real time MS-PCR, methylation sequencing, and pyrosequencing, as well as methylation microarray.

WHY CHOOSE THE METHYLAMP™ DNA MODIFICATION KIT?

• The fastest procedure available on the market, which can be completed within 1 hour and 55 minutes with consistent reaction conditions.
• Completely converts unmethylated cytosine into uracil: modified DNA > 99.98%.
• The lowest degradation of DNA in the modification process: more than 90% of DNA loss can be prevented.
• The lowest requirement of starting DNA for modification: only 50 pg or 20 cells.
• Extremely simple, reliable, and consistent modification conditions.

The Methylamp™ DNA Modification Kit contains all reagents required for bisulfite conversion on a DNA sample. DNA is chemically denatured to allow bisulfite reagent to react specifically with single-stranded DNA, thereby deaminating cytosine and creating a uracil residue. The unique DNA protection reagents contained in the modification buffer can prevent the chemical and thermophilic degradation of DNA in the bisulfite treatment. The non-toxic modified DNA capture buffer enables DNA to bind tightly to the column filter, thus DNA cleaning can be carried out on the column to effectively remove residual sodium bisulfite and salts. Modified DNA can then be eluted and stably stored at -20°C for up to 2 months.

High throughput version also available. [Cat. # P-1008]
Fast DNA Modification version using heating process also available. [Cat. # P-1010]

SCHEMATIC PROCEDURE:	COMPARATIVE OVERVIEW:
	The different amounts of DNA isolated from a serum sample were chemically modified using the Methylamp™ DNA Modification Kit. Real time PCR was performed by using a pair of primers and a probe designed to amplify both methylated and unmethylated alleles of β-actin.

KIT CONTENTS	40 samples P-1001-1	80 samples P-1001-2
R1 (DNA denature)	0.25 ml	0.5 ml
R2 (DNA modification)	4 vials	8 vials
R3 (DNA modification)	5 ml	10 ml
R4 (modified DNA capture)	14 ml	28 ml
R5 (modified DNA cleaning)	3 ml	6 ml
R6 (modified DNA elution)	1 ml	2 ml
F-spin column	40	80
F-collection tube	40	80
User guide	1	1

1. What is difference between one-step DNA modification and two-step DNA modification?
In the one-step DNA modification, DNA is denatured by heating, which allows DNA denaturation and bisulfite modification to be carried out simultaneously. One-step DNA modification is suitable for the labs which are equipped with thermal cycler and requires simple and fast procedures for DNA modification. In the two-step DNA modification, DNA is denatured chemically followed by bisulfite treatment. It is suitable for general DNA modification using DNA isloated from various sources. Because one-step DNA modification may increase DNA degradation, a higher starting DNA amount may be required for one-step DNA modification than for two-step DNA modifcation.

2. How much starting DNA is required for DNA modification with this kit?
The starting DNA required for DNA modification can be as low as 50 pg. The signal of the modified DNA can be detected with real-time PCR.

3. Why are only 90 minutes required for the actual DNA modification?
With our unique modification composition, 90 minutes is sufficient for more than 99% C-T conversion while DNA degradation is greatly prevented. We have observed that increase in modification time did not significantly increase C-T conversion, while yield of modified DNA was significantly reduced most likely due to increased DNA degradation.

4. Can the modified DNA with this kit be stored for a long time?
The modified DNA generated with this kit can be stored for 2 months at -20°C. The modified DNA could be stored for as long as 6 months at -80°C.

5. Can the kit be used for modifying DNA from formalin-fixed and paraffin-embedded (FFPE) samples?
DNA extracted from FFPE samples is often fragmented. The kit can be used for FFPE samples as the modification solution included in the kit contains DNA stabilizing reagents that avoid a further fragmentation of DNA caused in modification process.

6. Do I need to extract the DNA from blood first?
Yes. DNA isolation from blood is required and then DNA can be used for modification with the kit.

7. How much DNA can the columns actually handle?
The optimal range of DNA that can be used for each sample in each modification is 1ng to 1 µg. The column can maximally bind about 5 µg DNA.

8. Has anyone tried adding carrier DNA to their reactions when they use your kits?
We tested the carrier DNA together with the use of our kits and it only slightly increased yield of modified DNA.

9. When using this kit, can I start with cells or does the DNA have to be isolated first? Is there a product that efficiently recovers low amounts of DNA?
Yes. One can start with cells by using our whole cell bisulfite modification kit (P-1016) or start with isolated DNA by using P-1001 or P-1010. We provide a DNA isolation enhancer for recovering low amounts of DNA. However, as mentioned above, it may only slightly increase the recovery of DNA.

10. How should I store the kit, -20°C or just 4°C? ?
The kit can be stored at room temperature for at least 6 months or at 2-8°C for one year from shipping date.

[User Guide]* *Always use the actual User Guide that shipped with your product. Is the above file locked? You can also request user guides by emailing info@epigentek.com along with your contact information and institution name.

[Material Safety Data Sheet]

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