文献类型：期刊文献

英文题名：An efficient and cost-effective method for primer-induced nucleotide labeling for massive sequencing on next-generation sequencing platforms

作者：Guo, Junjie[1,3] Cheng, Tao[2] Xu, Han[1,3] Li, Yide[1,3] Zeng, Jie[1,3]

第一作者：郭俊杰;Guo, Junjie

通信作者：Li, YD[1];Zeng, J[1];Li, YD[2];Zeng, J[2]|[a000515db5a8e58e42817]李意德;

机构：[1]Chinese Acad Forestry, Res Inst Trop Forestry, Guangzhou 510520, Guangdong, Peoples R China;[2]Berry Genom Corp Ltd, Beijing 102200, Peoples R China;[3]Jianfengling Natl Key Field Res Stn Trop Forest E, Haikou 572500, Hainan, Peoples R China

年份：2019

卷号：9

期号：1

外文期刊名：SCIENTIFIC REPORTS

收录：;Scopus(收录号：2-s2.0-85062267192);WOS:【SCI-EXPANDED(收录号:WOS:000459897600059)】；

基金：This work was supported by National Nonprofit Institute Research Grant of Chinese Academy of Forestry, China (CAFYBB2011004-03) and National Natural Science Foundation of China (41201192). We thank Shuaibin Shang, Chunsheng Wang, and Mingxian Lin for their assistance on sample collection, Changhao Li for the helps in bioinformatics. We are also grateful to Professor Jianming Fu of the University of Kansas for editing the English language of this manuscript.

语种：英文

摘要：Next generation sequencing (NGS) technologies play a powerful role in the preparation of large DNA databases such as DNA barcoding since it can produce a large number of sequence reads. Here we demonstrate a primer-induced sample labeling method aiming at sequencing a large number of samples simultaneously on NGS platforms. The strategy is to label samples with a unique oligo attached to the 5'-ends of primers. As a case study, 894 unique pentanucleotide oligoes were attached to the 5'-ends of three pairs of primers (for amplifying ITS, matK and rbcL) to label 894 samples. All PCR products of three barcodes of 894 samples were mixed together and sequenced on a high throughput sequencing platform. The results showed that 87.02%, 89.15% and 95.53% of the samples were successfully sequenced for rbcL, matK and ITS, respectively. The mean ratio of label mismatches for the three barcodes was 5.68%, and a sequencing depth of 30x to 40x was enough to obtain reliable sequences. It is flexible to label any number of samples simply by adjusting the length of oligoes. This easy, reliable and cost efficient method is useful in sequencing a large number of samples for construction of reference libraries for DNA barcoding, population biology and community phylogenetics.