Briefly, B-ARISA polymerase chain reaction was performed in duplicate reactions under the conditions described by Borneman and Triplett [28] and modified according to Frossard et al. [15] and Purahong et al. [29]: the PCR mixture (20 μl) contained 1 μl DNA template (∼20 ng DNA template as determined by NanoDrop); 10 μM of primer 1406f (5′- TGYACACACCGCCCGT-3′) labeled with FAM at 5′-end and an unlabeled 23Sr primer (5′-GGGTTBCCCCATTC RG-3′); 4 μl FIREPol 5× Master Mix (Solis BioDyne, Tartu, Estonia); and water to 20 μl.