Illumina Genome Analyzer II sequencing; willow bud culture development
willow bud culture development
Extract 1
Axillary buds were isolated from willow stems (Salix purpurea genotype 844: Rothamsted National Willow Collection).Bud scales and stem epidermis removed under a dissecting microscope, and the isolated buds placed upright on plates containing root induction medium. After 4-6 weeks growth at 22-24 degrees C the roots were 1-2cm long and the plants were transferred to root induction medium  for another 10 days and then transplanted to soil and moved to a controlled environment cabinet at 20 degrees C /15 degrees C, 14h /10h  (day/night) and  330 micromolar m-2 s-1 light. The third stem internodes (counting from the basal stem) were collected at week 0 (before being transplanted to soil) and weeks 2, 4, and 6 (after transplantation to soil) and were either analysed immediately or frozen into liquid nitrogen. In order to obtain sufficient material for analyses, multiple internodes were pooled into samples (~70, ~50, ~20, 8 internodes at 0, 2, 4 and 6 weeks, respectively).  manufacturer's protocol for 110 bp SE reads