Sedimentary DNA reveals over 150 years of ecosystem change by human activities in Lake Chao, China

Feilong Lia , Xiaowei Zhanga,? , Yuwei Xieb , Jizhong Wangc,d a State Key Laboratory of Pollution Control & Resource Reuse, School of the Environment, Nanjing University, Nanjing 210023, PR China b Toxicology Centre, University of Saskatchewan, Saskatoon, Saskatchewan S7N 5B3, Canada c Guangzhou GRG Metrology & Test (Hefei) CO., LID, Hefei 230088, PR China d School of Environment and Energy Engineering, Anhui Jianzhu University, Hefei 230601, PR China

Universal eukaryotic primers (1380F: TCCCTGCCHTTTGTACACAC; 1510R: CCTTCYGCAGGTTCACCTAC) with attached unique 12-nt nucleotides (also known as tags) were used to amplify the 130 bp fragment of the V9 region of 18S rRNA genes (Amaral-Zettler et al., 2009). Universal bacterial primers (341F: ACCTACGGGRSGCWGCAG; 518R: GGTDTTACCGCGGCKGCTG) with attached unique tags were used to amplify the 180 bp fragment of the V3 region of 16 s rRNA genes (Klindworth et al., 2013; Muyzer et al., 1993). Primers were synthesized by Shanghai Generay Biotech Co., Ltd. Each sample was amplifified in triplicate PCR replicates and was subsequently pooled for the next procedure