The objective of this study was to investigate polymorphisms MASP2, TG5 and DQA1 genes using PCR-DNA sequencing in seventy Holstein, Brown Swiss dairy cows (35 each). Under complete aseptic condition, blood samples were collected from each animal into tubes containing disodium EDTA as an anticoagulant for DNA extraction. PCR carried out amplification 305-bp 545-bp TG5, 373-bp genes. assessment ...

Background: Ataxia with oculomotor apraxia type 1 (AOA1) shows early onset with autosomal recessive inheritance and is caused by a mutation in the aprataxin (APTX) gene encoding for the APTX protein. Methods: In this study, a 7-year-old girl born of a first-cousin consanguineous marriage was described with early-onset progressive ataxia and AOA, with increased cholesterol concentration and decr...

Microbial translocation (MT) from the gut is implicated in driving immune activation, increasing morbidity and mortality in HIV. We used bacterial 16S rDNA PCR, Sanger sequencing, and high-throughput sequencing to identify microbial DNA in the bloodstream of HIV-infected children in London, United Kingdom. Blood samples were collected from sequential children attending the HIV clinic at Great O...

478 | VOL.8 NO.6 | JUNE 2011 | nature methods and transcriptome ‘shotgun’ sequencing, next-generation DNA sequencing technologies are not readily applicable for identification of interacting pairs. The necessary pooling of PCR amplicons in the preparation of interacting sequence tags (ISTs) (Fig. 1a) would inevitably eliminate the association in each pair of DNA sequences coding for interacting...

OBJECTIVE To compare the real-time (RT), and qualitative (Q) polymerase chain reaction (PCR) assays for detection of Cytomegalovirus (CMV) DNA. METHODS The study took place in the Department of Microbiology, Erciyes University, Kayseri, and in Iontek Laboratory, Istanbul, Turkey, from August to December 2006. One hundred and seven clinical specimens from 67 patients were included in the study...

One of the major questions in microbial ecology is "who is there?" This question can be answered using various tools, but one of the long-lasting gold standards is to sequence 16S ribosomal RNA (rRNA) gene amplicons generated by domain-level PCR reactions amplifying from genomic DNA. Traditionally, this was performed by cloning and Sanger (capillary electrophoresis) sequencing of PCR amplicons....

An improved protocol is described for using lambda exonuclease to directly sequence PCR products. It is important not to execute PCR cycles beyond the plateau of amplification. The asymmetric PCR and double-stranded DNA sequencing by a snap-cooling procedure were also performed using the same DNA samples and primers. The improved method was the most reliable and produced the best results.