Navigation Links
Identification of Disease Causing Mutations in Phenylketonuria by Denaturing Gradient Gel Electrophoresis Using the DCode System

in compatible co-amplifications with comparable efficiencies, different exon PCR amplifications were combined. Moreover, to make sure that the resolving power to identify the exon specific mutations remains intact and that the DGGE patterns of the combined exons do not interfere with each other, the multiplex PCR amplifications have to be resolved subsequently on denaturing gradient gels. This resulted in a multiplex DGGE (MP-DGGE) of the PAH gene (Michiels et al. 1996).3 Examples of such an MP-DGGE are shown in Figure 2. The 13 different PAH exons are analyzed in 5 distinct multiplex PCR and DGGE patterns. Each exon shows distinct mutations, demonstrating that the resolving power of the DGGE gel remains intact. In addition, the DGGE patterns for the different exons do not overlap. Cycle sequencing of the DGGE DNA bands excised from the gel as described in the methods allows the identification of new mutations.

Several DNA preparation protocols have been tested: home made (see Methods) or commercially available kits such as Split Second from Boehringer Mannheim, Germany and DNA Easy-Prep from Lifecodes Corporation, USA, and can all be used in the MP-DGGE assay. All of them gave comparable results in the multiplex PCR/DGGE experiments.


Conclusion

In this report we demonstrate the use of powerful multiplex PCR amplification combined with DGGE analysis to rapidly identify mutations causing PKU. This MP-DGGE allows analysis of the complete PAH gene for three different individuals on one gel, whereas previously such analysis would take three gel runs. Moreover, the multiplex DGGE analysis is compatible with all the tested rapid DNA iso
'"/>

Source:


Page: All 1 2 3 4 5

Related biology technology :

1. Identification of Differentially Expressed Gene Products with the CastAway System*
2. An Optimal DNA Microarray Substrate for the Identification of Fetal and Somatic Liver Stem Cells of the Rat
3. Identification of Low-Level Oxidation Products in Calmodulin Using NanoSpray Ion Trap Mass Spectrometry
4. Detection and Identification of Phosphorylation Sites in Proteins Using LC/MS/MS with Neutral Fragment Loss Mapping
5. Protect RNA and Improve Target Cell Identification
6. Identification of Nonspecific Products Using Melt-Curve Analysis on the iCycler iQ Detection System, Rev A
7. Identification of cells in S phase using the Cell Proliferation Fluorescence Kit and IN Cell Analyzer 1000
8. Identification of Insecticides and Herbicides by LC/MS/MS Using the API 2000 LC/MS/MS System
9. Identification of Phase I and Phase II Metabolites of Buspirone on the Q TRAP LC/MS/MS System
10. ESR1 Gene Expression in Oncology and Metabolic Diseases Using the ASCENTATM System Target Validation and Identification of Novel Disease Indications
11. New PCR Primers for Human Diseases
Post Your Comments:
(Date:7/29/2015)... ... July 29, 2015 , ... Costello served as Lead Levee Design Engineering Manager ... Orange County, Texas area from future storm surge flooding, such as occurred in Hurricane ... levee alignment alternatives for providing protection, Costello also balanced socio-economic and environmental needs ...
(Date:7/28/2015)... ... , ... Strong Animals™ announced today that research conducted by South Dakota State ... ADSA-ASAS Joint Annual Meeting held July 12-16 in Orlando, FL showed feed additive Stay ... content. Also, feed containing Stay Strong™ for Dairy Cows took longer than 7.7 hours ...
(Date:7/28/2015)... , ... July 28, 2015 , ... ... than EUR 1.022 billion in 2013; while production volume reached close to 1.84 ... producing country with a share of 75.5% of the region’s total aluminium hydroxide ...
(Date:7/28/2015)... , July 28, 2015 Tauriga Sciences, ... diversified life sciences company, today announced that its stockholders ... of common stock of the Company from 1,000,000,000 to ... July 27, 2015 at the Law Offices of Nixon ... the Special Meeting, there were 480,655,929 shares of common ...
Breaking Biology Technology:Preventing Storm Surge Flooding in Houston 2Strong Animals™ Announces South Dakota State University Research Demonstrates Stay Strong™ for Dairy Cows Improves Performance and Feed Stability 2EU27 Aluminium Hydroxide Market Analysed and Forecast by BAC Reports in New Research Study Available at MarketPublishers.com 2EU27 Aluminium Hydroxide Market Analysed and Forecast by BAC Reports in New Research Study Available at MarketPublishers.com 3Tauriga Sciences, Inc. Stockholders Approve Increase In Authorized Common Stock 2Tauriga Sciences, Inc. Stockholders Approve Increase In Authorized Common Stock 3
... the information security firm Symantec released the 11th ... is Symantec's attempt to get out of the day-to-day and ... pretty ominous, according to Dave Cole, a director in the ... the game today is data theft, and the creation of ...
... Cagigal knows the missing link in many software evaluations, ... implementation. , ,Cagigal, chief information technology officer for ... on investment often neglect to factor in the ongoing ... the life of the application. , , David ...
... bioinformatics challenge, a free website has been developed by a ... and Children's Research Institute in Milwaukee to significantly improve diagnostic ... viruses. , ,The possibility of a pandemic has ... avian or bird flu virus, and they have been scrambling ...
Cached Biology Technology:Symantec: Cyber fraudsters look for low-hanging fruit 2Symantec: Cyber fraudsters look for low-hanging fruit 3Symantec: Cyber fraudsters look for low-hanging fruit 4Symantec: Cyber fraudsters look for low-hanging fruit 5Symantec: Cyber fraudsters look for low-hanging fruit 6Symantec: Cyber fraudsters look for low-hanging fruit 7The business case for software applications 2The business case for software applications 3Medical College team creates website for tracking flu 2Medical College team creates website for tracking flu 3
(Date:7/21/2015)... , July 21, 2015 Today, ZTE ... Already received as well as expected revenues in 2015 that relate ... FPC,s communicated revenue guidance of approximately 2,200 MSEK for 2015. ... is a leading smartphone manufacturer in China ... FPC1 02 5 for Axon , ...
(Date:7/13/2015)... , July 13, 2015 Synaptics ... of human interface solutions, today announced sampling of ... and display driver integration (TDDI) product targeting smartphones ... first to combine Synaptics , best-in-class touch ... display driver technology developed in the company,s Japan ...
(Date:7/9/2015)...  Unchained Labs just can,t get enough biologics ... Nano. Avid Nano designs, develops and manufactures highly ... Also today, Unchained Labs UNcaged the pUNk, ... protein sizing system. The pUNk is a high-performance ... size distribution, aggregation population and molecular weight. Its ...
Breaking Biology News(10 mins):FPC's Touch Fingerprint Sensor FPC1025 in ZTE's Smartphone Axon 2Synaptics Announces Sampling of Second Generation TDDI Solutions 2Synaptics Announces Sampling of Second Generation TDDI Solutions 3Unchained Labs just can't get enough, acquires Avid Nano, rocks out the new pUNk! 2
... is available in Spanish . , In ... research works that throw light on global warming are of ... atmospheric aerosol, a suspension of solid or liquid particles on ... cooling of the atmosphere. Juan Luis Guerrero Rascado ...
... of Singapore scientists, led by Christopher Wong, Ph.D., have ... of any influenza A virus, including H1NI, with just ... patients. The new method, which enables scientists ... sequence them within a day, was developed at the ...
... MDVariations in both milk feeding and in the weaning ... and they have independent influences on body composition in ... publication in The Endocrine Society,s Journal of Clinical ... Previous studies suggest that the early environment may be ...
Cached Biology News:A research work will be the reference to characterize the climatic impact of desert dust 2Rapid approach to identify influenza A virus mutations and drug resistance developed 2Breastfeeding duration and weaning diet may shape child's body composition 2
... C 4 H 10 O ... 154.25 PRODUCT SPECIFICATIONS Form: White crystalline ... to 97% Identity: By IR Melting ... 38C Oxidized DTT: less than or ...
... Feature Extraction Software reads and ... to prepare microarray data for ... * one 2-year license ... NFP/Academic accounts , * ...
... imported for data analysis , Automatic grid, subgrid ... template assignment , ... Extensive background and signal options to optimize, ... results , ...
... Stain combo offers these benefits: ... Offers a safer and ... Eliminates risks to yourself, the environment and your ... bp to >30 kb. UltraPure Agarose is a standard ...
Biology Products: