454 GS FLX+ sequencing; Comparative 454 pyrosequencing of transcripts to provide a resource for the study of embryogenesis of coconut (Cocos nucifera L.)
Comparative 454 pyrosequencing of transcripts to provide a resource for the study of embryogenesis of coconut (Cocos nucifera L.)
Immature embryos (9ME) and mature embryos (12ME) were sampled from 25 years old (in 2010) Sri Lanka Tall coconut palms growing at Bandirippuwa Estate, Lunuwila Sri Lanka. Total RNA was extracted using an RNeasy Plant Mini Kit (Qiagen, UK) using manufacturers protocol. cDNA libraries were constructed using SMART approach as described by Zhu et al. 2001 using the the oligonucleotides SMART Sfi1A : 5prime- AAG CAG TGG TAT CAA CGC AGA GTG GCC ATT ACG GCCrGrGrG-3', CDS-Sfi1B : AAG CAG TGG TAT CAA CGC AGA GTG GCC GAG GCG GCCd(T)20-3',  SMART PCR primer : 5'-AAG CAG TGG TAT CAA CGC AGA GT- 3'. The amplified cDNA PCR products were purified using QIAquick PCR Purification Kit (QIAGEN, CA) and a final  yield of 3 microgram was prepared for each tissue type.