![]() ![]() ![]() Typically, gene-specific oligonucleotides are used to amplify and tag a particular gene, for instance the ribosomal RNA gene (rDNA).īecause DNA ligation is relatively sequence-independent, the design of barcoded adapters using the ligated adapter strategy is straightforward. The second strategy, the topic of this paper, incorporates barcodes into oligonucleotides that are used to prime DNA synthesis, for instance, through polymerase chain reaction (PCR 5, 6 ). In one approach, barcoded, double-stranded adapters are ligated to target DNA through the activity of DNA ligase both shotgun genomic libraries and PCR amplicons can be tagged in this manner 7. Two strategies are in general use for barcoding samples in preparation for high-throughput sequencing. Life Sciences GS-FLX system 4 ) or by tagging genomic DNAs with unique, sample-specific sequences that serve as molecular barcodes 5, 6. High-throughput sequencing platforms now have the capacity to analyze multiple specimens in a single run. In many instances, project throughput can be accelerated by parallelization of sample processing, sequencing, and analysis steps. This paper describes the functions implemented by barcrawl and bartab and presents a proof-of-concept case study of both programs in which barcoded rRNA primers were designed and validated by high-throughput sequencing.Ĭonclusion Barcrawl and bartab can benefit researchers who are engaged in metagenomic projects that employ multiplexed specimen processing.Ĭoncomitant advances in automated DNA sequencing technology have greatly increased the scale of genomic and metagenomic studies 3. The program bartab can be used to deconvolute DNA sequence datasets produced by the use of multiple barcoded primers. Results Barcrawl is a software program that facilitates the design of barcoded primers, for multiplexed high-throughput sequencing. Primer 6 Statistical Software Software Program That Primer 6 Statistical Software Software Program That. ![]()
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