Pooled samples of Anastatica hierochuntica seedlings
Leaf tissue from 5 day-old in-vitro grown A. hierochuntica plants was harvested and snap-frozen in liquid nitrogen, and 100 mg of tissue was ground and used for RNA extraction using the RNeasy Mini Kit (Qiagen, 2001). RNA integrity was verified by denaturing formaldehyde agarose gel electrophoresis and the sample dried on GenTegra RNA transport tubes (Integen X, Pleasanton, CA, USA). RNA quality was confirmed on an Agilent Bioanalyzer at the Glasgow Polyomics Facility, University of Glasgow, UK. After DNase digestion, a cDNA library was prepared using the TruSeqTM RNA Sample Preparation kit (Illumina) followed by sequencing on an Illumina Genome Analyzer IIx (GAIIx).