NF-IR / Irradiated
Title ESTs from UV and gamma irradiated Medicago truncatula 21-day-old seedlings
Library name NF-IR, Irradiated
Library type EST, 5'-3' directional cDNA
Organism Medicago truncatula
Cultivar Jemalong (A17)
Tissue type Seedlings
Developmental stage 21 days old
Lab host E. coli strain SOLR (Stratagene)
Vector pBluescript SK-
Cloning site R1 (5') Eco RI
Cloning site R2 (3') Xho I
5' Adaptor sequence 5'-OH-AATTCGGCACGAGG-3'
3' Adaptor sequence 5'-(GA)10ACTAGTCTCGAG(T)17-3'
Description Seedlings were exposed either to 100 gy gamma or 0.5, 1, 5, or 10 kJ/m2 UV irradiation. Gamma-irradiated samples were harvested at 6, 12, 24 and 48 hours after treatment. UV-irradiated samples were harvested 24 hours post-treatment. cDNA was prepared from polyA+ enriched, pooled samples of equivalent amounts of total RNA from each sample. The cDNA was directionally ligated into the Uni-Zap XR vector (Stratagene) and packaged using the Gigapack III Gold packaging extracts. Phagemids containing cDNA inserts were in vivo excised from the recombinant Uni-ZAP XR vector using ExAssist helper phage and the E. coli strain XL1-Blue MRF' (Stratagene). Excised plasmids were plated using SOLR cells.
5' sequencing primer M13 Reverse
5' sequencing primer sequence 5'- TCA CAC AGG AAA CAG CTA TGA C -3'
3' sequencing primer N/A
3' sequencing primer sequence N/A
Other sequencing primer N/A
Other sequencing primer sequence N/A
Potential source of contaminant N/A
Additional comments  
Library developer Gregory D. May, Samuel Roberts Noble Foundation
Publication reference