GSM2507625: 6post; Ambystoma mexicanum; RNA-Seq
Samples were disrupted and homogenized using the TissueLyser (Qiagen), lysed in 600ul RLT Buffer and DTT (RNeasy Kit, Qiagen). Per sample one sterilized steel ball (Qiagen) was used in a 2 ml LoBind Tube (Eppendorf) at 30 Hz for 2 x 3 min. After homogenization and a short centrifugation the mix was transferred to a new tube and processed according to the RNeasy Kit (Qiagen) protocol for Animal Tissue (Handbook 06/2012). RNA sequencing libraries were prepared using mRNA Library Prep for Illumina RNA-Seq (Beckman Coulter) with an estimated fragment size of 300bp. SeraMag Poly-dT mRNA (GE health care life sciences) enrichment of mRNA was performed before sequencing 75bp single end reads on an Hiseq 2000 (Illumina).