Jack pine seedlings were provided by Mike Bendzsak (Forest First, SK) in January 2009. Seed originated from the 'Prince Albert West' seedlot in western Saskatchewan, and was initially grown at the PRT Armstrong Nursery field site. The seedlings were one year old when received at the University of Alberta. Seedlings were transplanted to 3.78 L plastic pots filled with Sunshine Mix #4 (Sun Gro Horticulture Canada TM) growing media. The plants were watered every two to three days in a growth chamber under the following conditions: 19oC (day/night) temperature, relative humidity of 20-35% and under full spectrum fluorescent lamps (200 micromol PAR) for a 15-h photoperiod. Wound treatment was applied to the stem of the previous year's growth with wounds made every 5 mm along two opposite sides of the stem. The full length of the stem was mechanically wounded in April 2009 by making horizontal cuts with a razor blade. Samples (xylem and bark) were collected along the wounded section of stem at 1, 2, 4 and 8 days after wounding. Xylem and bark of four replicates for each time point were collected. Stem sections were cut and xylem and bark were separated manually and immediately frozen independently in liquid nitrogen and stored at -80oC until RNA extraction. A normalized, full-length enriched, directionally cloned library was prepared by Evrogen (Moscow, Russia) with the Creator SMART cDNA Library Construction Kit (Clontech, Mountain View, CA, USA) and the TRIMMER-DIRECT cDNA Normalization Kit (Evrogen). Total RNA was individually extracted from each bark tissue sample using the large scale RNA extraction method described in Chang, S., et al. (1993) Plant Mol. Biol. Reporter 11(2):113-116 and DNase I treated (New England Biolabs). Total RNA from one replicate for each time point was pooled and 17 micrograms of total RNA was sent to Evrogen. First strand cDNA was prepared from 300 nanograms of total RNA, PowerScript Reverse Transcriptase (Clontech), CDS-3M primer (Evrogen), and the SMART IV Oligonucleotide (Clontech). Second strand cDNA was prepared by LD-PCR with Advantage 2 Polymerize mix (Clontech). This cDNA was then normalized with duplex-specific nuclease and amplified following the TRIMMER-DIRECT protocol, digested with SfiI, size fractionated, and cDNA larger than approx. 1000 bp was ligated into SfiI-digested pDNR-LIB (Clontech). The ligation mixture was transformed into ElectroMAX DH10B T1 Phage-Resistant electro-competent cells (Invitrogen). Titre of the primary library (1 mL) was approx. 6.3x10^5 cfu/mL and was not amplified.