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PDF S7806 Data sheet ( Hoja de datos )
| Número de pieza | S7806 | |
| Descripción | CpG WIZ Prader-Willi/Angelman Amplification Kit | |
| Fabricantes | ETC | |
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CpG WIZ™ p16 Amplification Kit
S7800
CpG WIZ™ p15 Amplification Kit
S7802
CpG WIZ™ E-cadherin
Amplification Kit
S7804
CpG WIZ™ Prader-Willi/Angelman
Amplification Kit
S7806
FOR RESEARCH USE ONLY
Not for use in diagnostic procedures
USA & Canada
Phone: +1(800) 437-7500 • Fax: +1 (909) 676-9209 • Europe +44 (0) 23 8026 2233
Australia +61 3 9839 2000 • Germany +49-6192-207300 • ISO Registered Worldwide
1 page  
I. INTRODUCTION
Using This Manual
Please read the entire instruction manual prior to using CpG WIZ™
Amplification Kits. Note that in the Procedures and Troubleshooting sections,
instructions are separate for the CpG WIZ™ Prader-Willi/ Angelman
Amplification Kit (S7806). Should additional questions arise, assistance is
available from Chemicon Technical Service at [email protected] or
(800) 437-7500.
Background
Methylation of cytosines located 5' to guanosine is known to have a profound
effect on the expression of several eukaryotic genes (1). In normal cells,
methylation occurs predominantly in CG-poor regions, while CG-rich areas,
called CpG islands, remain unmethylated. The exception is extensive
methylation of CpG islands associated with transcriptional inactivation of
regulatory regions of imprinted genes (2, 3) such as those associated with
Prader-Willi/Angelman Syndrome (4) and genes on the inactive X-chromosome
of females (5, 6). Aberrant methylation of normally unmethylated CpG islands
has been documented as a relatively frequent event in immortalized and
transformed cells (7) and has been associated with transcriptional inactivation of
defined tumor suppressor genes in human cancers (8, 9). E-cadherin, p16, and
p15 are examples of genes that exhibit characteristic hypermethylation.
Previously developed methods to determine the methylation status of cytosine
include digestion with methylation sensitive restriction enzymes and genomic
DNA sequencing. Both techniques have limitations: restriction enzymes can
only detect methylation sites within their recognition sequence and sequencing
is time consuming. Increasing the detection sensitivity of CpG island
methylation has the potential to define tumor suppressor gene function and
provides a new strategy for early tumor detection.
Methylation-specific PCR (MSP) is a new technology for sensitive detection of
abnormal gene methylation utilizing small amounts of DNA (10). This process
employs an initial bisulfite reaction to modify the DNA, followed by PCR
amplification with specific primers designed to distinguish methylated from
unmethylated DNA. The CpGenome™ DNA Modification Kit (S7820) contains
the reagents necessary to perform the initial bisulfite reactions, while CpG
WIZ™ Amplification Kits contain the reagents required for the PCR
amplification reactions.
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5 Page 
III. PROTOCOLS
CpG WIZ™ p16 (S7800), p15 (S7802) and E-cadherin (S7804)
Amplification Kits
Experimental Design
Primer Sets
CpG WIZ™ Amplification Kits contain primers that can be used for analysis of
DNA samples by MSP. However, the samples must first undergo bisulfite
modification prior to PCR amplification. CpGenome™ DNA Modification Kit,
S7820, contains the reagents necessary to perform the modification. Chemical
modification creates the sequence differences between the methylated and
unmethylated DNA. The primer sets in the kit are engineered to anneal to the
DNA, based upon the sequence differences.
U Primer Set will anneal to unmethylated DNA that has undergone a chemical
modification
M Primer Set will anneal to methylated DNA that has undergone a chemical
modification
W Primer Set serves as a control for the efficiency of chemical modification. It
will anneal to any DNA (unmethylated or methylated) that has NOT undergone
chemical modification, hence, the "wild type", or W.
Data interpretation can still proceed in the case of incomplete chemical
modification (up to 50%).
Amplification Regions
The amplified region is defined as the sequence between the 3' nucleotide of the
sense primer and the complement of the 3' nucleotide of the anti-sense primer
for each gene promoter. The nucleotide numbering systems are those used in the
GenBank submissions identified by the following accession numbers: p16,
X94154; p15, S75756; E-cadherin, L34545.
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11 Page | ||
| Páginas | Total 30 Páginas | |
| PDF Descargar | [ Datasheet S7806.PDF ] | |
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| S7804 | CpG WIZ Prader-Willi/Angelman Amplification Kit | ETC |
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