Sample collection and DNA extractio

Sample collection and DNA extraction
To answer the above questions, we sequenced the mitochon- drial cytochrome b gene from 448 individuals. The study sample
represents 16 adults of L reevesii collected between 2007 and 2008 from each of 28 sampling localities almost covering the spe- cies’ distribution range from Chinese mainland (Guangdong and Guangxi) to Hainan (the largest island second to Taiwan in China) and Vietnam (Fig. 1). During our collecting trip to the coastal re- gion of Vietnam and Guangxi (a province of China), we did not find any L. reevesii in the north of Vietnam and in the west of Guangxi, close to Vietnam. This is the region where the microcontinental blocks of Southeast Asia and the megacontinent of Asia contacted (Fig. 1), currently having no habitat suitable for L. reevesii.
The most distal 15 mm of the tail tip of each lizard was excised using a sterilized scalpel. Individual lizards were released at their site of capture after tissue sampling. Tissue samples were pre- served in 95% ethanol before they were deposited at Hangzhou Normal University and assigned voucher numbers identified by locality-haplotype numbers. Total DNA was extracted using stan- dard phenol–chloroform methods (Sambrook et al., 1989), and stored at