NF-PL / Phosphate-Starved Leaves
Title ESTs from phosphate-starved Medicago truncatula leaves
Library name NF-PL, Phosphate-starved leaves
Library type EST, 5'-3' directional cDNA
Organism Medicago truncatula
Cultivar Jemalong (A17)
Tissue type Aerial tissues (leaves and stems)
Developmental stage Mature plant
Lab host E. coli 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 At the trifoliate stage, M. truncatula plants were transplanted to phosphate-free sand and grown for a further 30 days. During this 30 day period, the plants were fertilized twice weekly with 1/2 Hoaglands solution containing only 20 uM potassium phosphate. RNA was prepared from above ground tissues. 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 Jinyuan Liu, Maria J. Harrison, Samuel Roberts Noble Foundation
Publication reference