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Timothy b schmit torrent4/17/2023 By using high-throughput machines (PCR and NGS), labeling PCR, and the Cotu method, it is possible to significantly reduce the cost and labor investments for DNA barcoding. Step-by-step instructions to our method are provided. ![]() We developed a new “Cotu” method to create consensus sequences from NGS reads for longer output sequences and more reliable bases than the other three methods. After comparing the sequencing depths, read lengths, base qualities, and base accuracies, we conclude that Illumina Hiseq2500 PE250 run is suitable for conventional DNA barcoding. ![]() We sequenced six conventional DNA barcode fragments (ITS1, ITS2, matK1, matK2, rbcL1, and rbcL2) of 380 flowering plants on next-generation sequencing (NGS) platforms (Illumina Hiseq 2500 and Ion Torrent S5) and the Sanger sequencing platform. Here, we present our solutions to these challenges. Unfortunately, there are experimental and data processing challenges to construct such a library within a short time. Successful application of this technology is dependent on the availability of reference database of high species coverage. DNA barcoding has become one of the most important techniques in plant species identification.
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