RNA-Seq of Bufo bufo : pool of 9 metamorphs head
Juveniles were individually sampled and immediately placed in liquid nitrogen and then stored at -80C until molecular analyses. Only junevile's head was kept. Therefore, total RNA was extracted from individual head by adding TRI Reagent and homogenizing tissue on ice with a tissue lyser. Then RNA was treated with Turbo DNase enzyme and assayed by flurorescence with Qubit fluorometer. mRNA was isolated and pooled to allow the construction of one cDNA library using TruSeq RNA Sample Prep Kit v2 -Set A (ILLUMINA, RS-122-2001), which was sequenced with an Illumina HiSeq4000.