Illumina Genome Analyzer IIx sequencing; Musa acuminata accession DH-Pahang leaf pieces mock inoculated
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.
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 108-base length read chemistry in a single flow cell on the Illumina GA IIx (Illumina, USA).