Go Back   SEQanswers > Literature Watch

Similar Threads
Thread Thread Starter Forum Replies Last Post
PubMed: Genome-wide genetic marker discovery and genotyping using next-generation seq Newsbot! Literature Watch 0 08-30-2011 03:03 AM
PubMed: Genome structural variation discovery and genotyping. Newsbot! Literature Watch 1 04-20-2011 01:22 AM
PubMed: The JCVI standard operating procedure for annotating prokaryotic metagenomic Newsbot! Literature Watch 0 02-10-2011 07:10 AM
PubMed: Next generation transcriptomes for next generation genomes using est2assembly Newsbot! Literature Watch 0 12-26-2009 03:18 AM
PubMed: Pyrosequencing for SNP genotyping. Newsbot! Literature Watch 0 09-22-2009 03:00 AM

Thread Tools
Old 12-08-2010, 03:30 AM   #1
RSS Posting Maniac

Join Date: Feb 2008
Posts: 1,443
Default PubMed: Overcoming methodical limits of standard RHD genotyping by next-generation se

Syndicated from PubMed RSS Feeds

Related Articles Overcoming methodical limits of standard RHD genotyping by next-generation sequencing.

Vox Sang. 2010 Dec 7;

Authors: Stabentheiner S, Danzer M, Niklas N, Atzmüller S, Pröll J, Hackl C, Polin H, Hofer K, Gabriel C

Background and objectives Molecular variations of the RHD gene may result in the reduced expression of the D antigen and altered Rh phenotypes. In many occasions, they cannot be typed reliably by standard serological methods. Sequence-based typing is the gold standard to determine rare and unknown RHD genotypes. For this pilot study, sequence-based typing by standard Sanger sequencing was compared to a newly established next-generation sequencing approach based on pyrosequencing. Materials and methods Twenty-six DNA samples were selected after primary serological testing exhibiting a weak reaction in Rh phenotype. Parallel sequence analysis of the complete coding sequence including adjacent intronic sequences allowed a comparison of the methodical potency in mutation detection of Sanger with next-generation sequencing. Results Sanger sequencing revealed 39 RHD polymorphisms in 21 of 26 samples in the RHD coding region, while pyrosequencing detected all but two alterations resulting in a concordance rate of 94·9% and clearly revealed a heterozygous compound mutation in one sample with RHD? and Weak D type 4 alleles. The resolution of cis/trans linkage of polymorphisms and exact characterization of a 37 bp duplication was achieved by next-generation sequencing. Conclusion Our data suggest that next-generation sequencing offers a new development for high-throughput and clonal sequencing for molecular RHD genotyping. However, further attempts in the methodical set-up have to be undertaken prior to validation and introduction as a routine service.

PMID: 21133932 [PubMed - as supplied by publisher]

Newsbot! is offline   Reply With Quote

Thread Tools

Posting Rules
You may not post new threads
You may not post replies
You may not post attachments
You may not edit your posts

BB code is On
Smilies are On
[IMG] code is On
HTML code is Off

All times are GMT -8. The time now is 02:41 AM.

Powered by vBulletin® Version 3.8.9
Copyright ©2000 - 2021, vBulletin Solutions, Inc.
Single Sign On provided by vBSSO