title: Illumina Genome Analyzer IIx sequencing; Musa acuminata accession DH-Pahang leaf pieces inoculated with Mycosphaerella fijiensis strain CIRAD139A
design: Inoculations with M. fijiensis strain CIRAD139A were performed in controlled conditions for on leaf pieces (25 cm2) from three banana accessions with a differential response to M. fijiensis. Leaf fragments were spray-inoculated either with water (mock inoculation) or a suspension of mycelial fragments. RNA extraction was performed at 10 and 16 days post-inoculation (dpi). Each RNA extraction pooled half of a leaf fragment (12.5 cm2) from 2 different plants of the same accession. RNA from 10 dpi and 16 dpi leaves was pooled for cDNA library construction.
library_construction_protocol: cDNA libraries were constructed using the SMARTer PCR cDNA Synthesis Kit (Clontech, Saint-Quentin-en-Yvelines, France) according to the manufacturer's instructions, using 1 ug of DNase-treated total RNA. The cDNAs were sonicated to a 150- to 600-bp size range using the S2 Covaris instrument (Covaris, Inc., USA). Single end libraries were prepared using standard Illumina protocols (Illumina DNA sample kit). Each library was sequenced using the 76-base length read chemistry in a single flow cell on the Illumina GA IIx (Illumina, USA).