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DNA甲基化修饰试剂盒
产品名称: DNA甲基化修饰试剂盒

产品货号: P-1001

产品产地: Epigentek

产品规格: 40/80 

产品价格:  1800/3180

产 品 说 明:

The Methylamp#8482; 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#8482; 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#8482; 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.

Principle & Procedure


The Methylamp#8482; 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 clean DNA 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#8482; DNA Modification Kit. Real time PCR was performed by using a pair of primers and a probe designed to amplify both

Product Components

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

Frequently Asked Q's


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.

References & Citations

Mizoguchi, Yoko et al. (Jan 2007). Steroid-Dependent ACTH-Produced Thymic Carcinoid: Regulation of POMC Gene Expression by Cortisol via Methylation of Its Promoter Region. Hormone Research 67(5): 257-62. PubMed Abstract

Li, Kay Ka Wai et al. (Dec 2006). EMP3 overexpression is associated with oligodendroglial tumors retaining chromosome arms 1p and 19q. International Journal of Cancer 120(4): 947-50. PubMed Abstract

Tong, Yu K et al. (Oct 2006). Noninvasive prenatal detection of fetal trisomy 18 by epigenetic allelic ratio analysis in maternal plasma: Theoretical and empirical considerations. Clin. Chem. 52(12): 2182-3. PubMed Abstract

Cheng, Suk Hang et al. (Oct 2006). 4q loss is potentially an important genetic event in MM tumorigenesis: identification of a tumor suppressor gene regulated by promoter methylation at 4q13.3, platelet factor 4. Blood DOI 10.1182: blood-2006-04-01877 PubMed Abstract

[User Guide]
Is the 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]

www.epigentek.com/catalog/product_info.php?cPath=21&products_id=28&osCsid=ed2c2cbe6f502268629975b6d51d14fc


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