**3.1 Fish sampling, DNA isolation, and quantification and microsatellite amplification**

Sampling of Silver Carp (*H. molitrix*) was done from five selective sites of River Chenab, namely Trimmu Barrage, Marala Headworks, Khanki Headworks, Qadirabad Headworks, and *Chiniot* Bridge (**Table 1**) and in total 175 samples were collected (35 samples from each site). By following Sambrook and Russell [43], DNA isolation protocol having slight modifications, DNA was extracted from dorsal muscle tissues. By using scissors, spines and scales were removed as much as possible from frozen tissues. Standard proteinase-K and phenol/chloroform method was used for isolating DNA. Genomic DNA isolation consisted of following steps:


### **Tissue lysis buffer:**

