Transcriptome of Calotropis procera
Library  preparation  was  done following Illumina  TruSeq  RNA  library  protocol  outlined in  “TruSeq  RNA    Sample  Preparation  Guide” .Briefly, 1ug of Total RNA by qubit was enriched for PolyA  using  RNA  Purification  Beads  provided  with  the  kit,  enriched  RNA  was  fragmented  for  4   minutes  at  elevated  temperature  (94  o  C)  in  the  presence  of  divalent  cations  and  reverse   transcribed  with  Superscript  III  Reverse  transcriptase  by  priming  with  Random  Hexamers.   Second strand cDNA was synthesized in the presence of DNA Polymerase I and RnaseH. The   cDNA  was  cleaned  up  using  AgencourtAmpure  XP  SPRI  beads  (Beckman  Coulter).  Illumina   Adapters were ligated to the cDNA molecules after end repair and addition of  “A”-  base. SPRI   cleanup  was  performed  after  ligation.  The  library  was  amplified  using  8  cycles  of  PCR  for  enrichment  of  adapter  ligated  fragments.