title: Chicken and turkey sausage
design: Dual indexed amplicons. The first amplification was performed primer pair which contained an additional Illumina TruSeq adaptor on 1-10 ng of DNA extract (total volume 1l). For the second amplification, we used 1 l of each amplicon obtained in the first PCR, and these amplicons were separately amplified in a 20 l reaction volume using standard i7- and i5- sequencing adaptors. For both the first and second PCR, we used 15 pmol of each forward and reverse primer in 20 L volume of 1x MyTaq buffer containing 1.5 units MyTaq DNA polymerase (Bioline GmbH, Luckenwalde, Germany) and 2 l of BioStabII PCR Enhancer (Sigma-Aldrich Co.). PCRs were run for 30 cycles. Setting for first amplification: 1 min 96C pre-denaturation followed by 15 s at 96C denaturation, 30 s at 58C annealing and 90 s at 70C extension and a final hold at 8C. Second amplification was as first amplification with modified annealing temperature, i.e. 3 cycles at 50C followed by 7 cycles at 58C