| Title |
ESTs from elicited Medicago truncatula cell cultures |
| Library name |
NF-EC, Elicited cell culture |
| Library type |
EST, 5'-3' directional cDNA |
| Organism |
Medicago truncatula |
| Cultivar |
Jemalong (A17) |
| Tissue type |
Cell cultures derived from root tissues |
| Developmental stage |
Cell suspensions were subcultured every 14 days. Cells were induced six days after subculture. |
| 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 |
Cells were induced with yeast cell wall extract at a final concentration of 50ug glucose equivalents per ml. Samples were taken at 0.5, 1, 12 and 24 hours after induction. Equal amounts of RNA from each time point were pooled. 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 |
Richard A. Dixon, Samuel Roberts Noble Foundation |
| Publication reference |
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