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Sequences with pairwise BLASTN matches of Lenalidomide species were designed based on alignments of the genes (Additional file 9) and recommendations in the Phusion HotStart polymerase instruction manual. PCR reactions were performed in 1?Phusion HF buffer with 1.5 mM MgCl2, PubMed ID:https://www.ncbi.nlm.nih.gov/pubmed/17139194 200 M dNTPs, 0.5 M of forward andRoxstr -Lindquist et al. BMC Genomics 2010, 11:258 http://www.biomedcentral.com/1471-2164/11/Page 12 ofreverse primers, 40 ng S. barkhanus or S. salmonicida genomic DNA and 0.8 U Phusion HS DNA polymerase (Finnzymes) in a total volume of 40 l. The.