您所在的位置:首页 > DNA甲基化修饰试剂盒
DNA甲基化修饰试剂盒

【产品中文名称】

 DNA甲基化修饰试剂盒

【产品产地】

 Epigentek

 English Name

 Methylamp DNA Modification Kit

产品货号

 P-1001

 产品 价格/规格

 40/1700,80/3180

 【产品详细说明】

    

Methylamp DNA Modification Kit

Product Overview



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.

Principle & Procedure


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 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™ 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.

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.

User Guide & MSDS

[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]

References & Citations


Liu, T et al. (January 2009). Increased {gamma}-tubulin expression and P16INK4A promoter methylation occur together in preinvasive lesions and carcinomas of the breast. Annals of Oncology.
Epub ahead of print. PubMed Abstract

Ramírez, N et al. (November 2008). Epigenetic events in normal colonic mucosa surrounding colorectal cancer lesions. European Journal of Cancer.
44(17): 2689-95. PubMed Abstract

Karakoula, Katherine et al. (Nov 2008). Real-time quantitative PCR analysis of pediatric ependymomas identifies novel candidate genes including TPR at 1q25 and CHIBBY at 22q12-q13. Genes Chromosomes Cancer. 47(11): 1005-22. PubMed Abstract

Cheng, CK et al. (October 2008). Transcriptional repression of the RUNX3/AML2 gene by the t(8;21) and inv(16) fusion proteins in acute myeloid leukemia. Blood.
112(8): 3391-402. PubMed Abstract

Wiley, Christopher D. et al. (Oct 2008). Patterns of Hybrid Loss of Imprinting Reveal Tissue- and Cluster-Specific Regulation. PLoS ONE. 3(10): e3572. PubMed Abstract

Sasai, Ken et al. (Aug 2008). Careful Exclusion of Non-neoplastic Brain Components is Required for an Appropriate Evaluation of O6-methylguanine-DNA Methyltransferase Status in Glioma: Relationship Between Immunohistochemistry and Methylation Analysis. Am J Surg Pathol.
32(8): 1220-7. PubMed Abstract

Sanosaka, T et al. (August 2008). Identification of genes that restrict astrocyte differentiation of midgestational neural precursor cells. Neuroscience.
155(3): 780-8. PubMed Abstract

Kiang, Xing-Wei et al. (Jun 2008). Loss of methylation imprint of Snrpn in postovulatory aging mouse oocyte. Biochem Biophys Res Commun. 371(1): 16-21. PubMed Abstract

Torrisani, Jérôme et al. (Jun 2008). Identification of an upstream promoter of the human somatostatin receptor, hSSTR2, which is controlled by epigenetic modifications. Endocrinology 149(6): 3137-47. PubMed Abstract

Shirohzu, Hisao et al. (May 2008). Methylation of a conserved intronic CpG island of mouse SF-1 is associated with cell-specific expression of SF-1 in a culture system but not with tissue-specific expression. Biochemical and Biophysical Research Communications.
369(3): 862-7. PubMed Abstract

Britschgi, Christian et al. (Apr 2008). HIC1 tumour suppressor gene is suppressed in acute myeloid leukaemia and induced during granulocytic differentiation. Br J Haematol. 141(2): 179-87. PubMed Abstract

Ushmorov A, Leithäuser F et al. (Mar 2008). ABF-1 is frequently silenced by promoter methylation in follicular lymphoma, diffuse large B-cell lymphoma and Burkitt's lymphoma. Leukemia Epub Ahead of Print. PubMed Abstract

Olsson, Mats et al. (Jul 18 2007). Regulation and expression of human CYP7B1 in prostate: Overexpression of CYP7B1 during progression of prostatic adenocarcinoma. Prostate 67(13): 1439-1446. PubMed Abstract

Tokita, Tomoko et al. (Mar 2007). Methylation status of the SOCS3 gene in human malignant melanomas. International Journal of Oncology 30(3): 689-94. PubMed Abstract

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


pyright 2004-2009 www.bioso.cn 金佰利娱乐有限公司版权所有 订货热线:4008811876
电话:010-62917543 87175666 87175777 传真:010-62913373 邮箱:bioso@yahoo.cn 技术:biosou@yahoo.cn 网站统计