Navigation Links
Detection of Mutations in the CYP21 Gene Using the DCode System

Gita Ohlsson and Marianne Schwartz, Department of Clinical Genetics, Juliane Marie Center, University Hospital, Rigshospitalet, Copenhagen, Denmark


Introduction
Congenital adrenal hyperplasia (CAH) due to 21-hydroxylase deficiency is caused by mutations in the gene CYP21 encoding the enzyme steroid 21-hydroxylase. In addition to deletions, approximately 20 different point mutations have been reported14 and novel mutations are still detected.58 This makes genetic diagnosis as well as carrier detection of 21-hydroxylase deficiency a complicated matter. We have used the denaturing gradient gel electrophoresis method (DGGE) using the DCode universal mutation detection system to detect mutations in the coding sequence and intron-exon junctions of CYP21.9, 10


Materials and Methods
Samples
DNA from healthy individuals and patients with 21-hydroxylase deficiency was isolated from peripheral blood lymphocytes using the salting-out method by Miller et al.11

Polymerase Chain Reaction (PCR)
For amplification of CYP21 and selection against CYP21P, PCR was performed according to Wedell.12 The resulting two PCR products were purified and subsequently used as template for the amplification of DNA fragments for DGGE. The PCR reactions were performed with GC-clamped primers. The GC-density of almost all CYP21 exons are very high requiring long GC-clamps to obtain a single melting domain of the sequence of interest.

Site-Directed Mutagenesis
In order to obtain mutant control samples for all exons it was necessary to introduce mutations i
'"/>

Source:


Page: All 1 2 3 4 5

Related biology technology :

1. Simple, Sensitive, and Rapid Detection of FLAG -Tagged Fusion Proteins
2. A New PCR-based Mycoplasma Detection Method
3. HSVision Molecular Beacon Detection Module Rapidly Detects Herpes Simplex Virus DNA
4. Detection and Identification of Phosphorylation Sites in Proteins Using LC/MS/MS with Neutral Fragment Loss Mapping
5. Detection of mRNAs on Cryosections of the Cardiovascular System Using DIG-Labeled RNA Probes
6. Gene Expression Arrays: Highly Sensitive Detection of Expression Patterns with Improved Tools for Target Amplification
7. The DIG System Nonradioactive and Highly Sensitive Detection of Nucleic Acids
8. Quantitative Measurement of Cell Proliferation Using the BrdU ELISA: A Comparison Between Colorimetric and Chemiluminescent Detection
9. Quantification of Nucleosomes in Serum by the Cell Death Detection ELISAplus
10. A Further Step in Understanding Apoptosis Direct Detection of PARP Cleavage
11. In Situ Cell Death Detection Kit
Post Your Comments:
*Name:
*Comment:
*Email:
TAG: Detection Mutations the CYP21 Gene Using the DCode System

(Date:8/1/2014)... (PRWEB) August 01, 2014 RURO, ... Solutions provider, today officially announces the opening of ... Boji Science Park, Baoshan District in Shanghai, PRC, ... is under the management of Mr. Shen Gang, ... In the past three years, RURO’s has installed ...
(Date:8/1/2014)... York , August 1, 2014 ... Transparency Market Research "Biophotonics Market - Global Industry Analysis, ... the market was valued at USD 25.16 billion in ... by 2020, growing at a CAGR of 10.4% from ... Market Report at  http://www.transparencymarketresearch.com/biophotonics-market.html Increasing demand ...
(Date:8/1/2014)... , Aug. 1, 2014 Glencoe ... launch of "Data InPress" ( http://glencoesoftware.com/data-inpress.html ), ... integrating multidimensional image data into on-line publications.  ... and Video Injection Service. Photo ... http://photos.prnewswire.com/prnh/20140731/132409 The ...
(Date:7/31/2014)... Nearly 20,000 medical professionals and healthcare leaders ... Clinical Lab Expo in Chicago ... diagnostic research and technology that will advance medicine and ... of Wednesday, July 30, more than 19,500 attendees had ... Lab Expo, with more than 10,000 of these attendees ...
Breaking Biology Technology:RURO Opens Office in Shanghai, China 2Global Biophotonics Market is Expected to Reach USD 50.20 Billion in 2020: Transparency Market Research 2Global Biophotonics Market is Expected to Reach USD 50.20 Billion in 2020: Transparency Market Research 3Global Biophotonics Market is Expected to Reach USD 50.20 Billion in 2020: Transparency Market Research 4Glencoe Software Launches Data InPress 2Glencoe Software Launches Data InPress 3Research on Digital Health, Alzheimer's, Ebola Draws Nearly 20,000 Attendees to 2014 AACC Annual Meeting & Clinical Lab Expo 2Research on Digital Health, Alzheimer's, Ebola Draws Nearly 20,000 Attendees to 2014 AACC Annual Meeting & Clinical Lab Expo 3
... supplier of automatic identification and data collection (AIDC) software, ... their NiceLabel software. , ,The new versions of NL ... identification (RFID) tags, as well as an improved user ... programs such as Microsoft Access, Windows Event Log and ...
... Paragon Development Systems (PDS), an Oconomowoc-based IT services ... with Cisco Systems . Starting July 1, the ... services and communications markets. , ,PDS recently received Specialized ... areas in which the companies will partner; IP Communications, ...
... lawsuit on behalf of Wisconsin consumers against Microsoft can ... of last week's state Supreme Court ruling . ... seeks a settlement fund for all Wisconsin consumers for ... is that Wisconsin consumers have paid inflated prices for ...
Cached Biology Technology:Wisconsin Supreme Court rules against Microsoft 2
(Date:8/1/2014)... fossil record about 200,000 years ago, but it was ... advanced tools became widespread. , A new study ... finds that human skulls changed in ways that ... same time that culture was blossoming. , "The modern ... cultural exchange probably came at the same time that ...
(Date:8/1/2014)... Knowing what causes them at the molecular level is ... have created the most detailed map to date of ... blinding diseases, such as age-related macular degeneration. The high-resolution ... which supplies blood and oxygen to the outer retina, ... abundance of proteins in different areas of the choroid, ...
(Date:8/1/2014)... One of the most diverse families in the ocean ... lucinids) -- originated more than 400 million years ago ... like those of its modern members. This Geology ... Hawaii, published online on 25 July 2014, tracks the ... relationships. , At is origin, the Lucinidae family remained ...
Breaking Biology News(10 mins):Society bloomed with gentler personalities and more feminine faces 2A map for eye disease 2Symbiotic survival 2Symbiotic survival 3Symbiotic survival 4Symbiotic survival 5Symbiotic survival 6Symbiotic survival 7Symbiotic survival 8Symbiotic survival 9Symbiotic survival 10Symbiotic survival 11Symbiotic survival 12Symbiotic survival 13Symbiotic survival 14
... School of Medicine have found evidence that a hormone ... functions of spatial learning and memory development, and further ... stomach. , Published in the February 19 online issue ... institutes, the study showed that the hormone ghrelin, produced ...
... cartilage offers the first hope of replacing the ... The technique, developed at Rice University's Musculoskeletal Bioengineering ... the journal Tissue Engineering. , "This has significant ... for the first time, about the potential treatment ...
... led by a University of Minnesota researcher has found a ... kinds and sizes and that may also offer a key ... be known precisely in order to construct valid models of ... occur in both plants and animals through the process of ...
Cached Biology News:Learning and memory stimulated by gut hormone 2Rice bioengineers pioneer techniques for knee repair 2Of mice, men, trees and the global carbon cycle 2Of mice, men, trees and the global carbon cycle 3
I-Ab MHC Class II...
... mm Culture Dishes BD BioCoat Collagen I ... a uniform application of rat tail collagen type ... ,Surface : ECM/attachment factors ,Surface ... : 60 mm x 15 mm ...
The piston kit includes 2 pistons to service one F10 pump of a BioLogic DuoFlow workstation....
BD BioCoat BD MatrigelTM Matrix 100 mm Culture Dishes for hepatocytes, tissue-culture treated polystyrene with a uniform application of MATRIGEL Matrix....
Biology Products: