| Title |
ESTs from Phoma medicaginis-inoculated leaves of Medicago truncatula |
| Library name |
Phoma-infected (NF-PH) |
| Library type |
EST, 5'-3' directional cDNA |
| Organism |
Medicago truncatula |
| Cultivar |
Jemalong (A17) |
| Tissue type |
Leaf |
| Developmental stage |
Pathogen-induced, young trifoliate |
| 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 |
Young trifoliate leaves of Medicago truncatula were excised and dip-inoculated in a spore suspension of Phoma medicaginis, and incubated in humid dishes. Pools of leaves were harvested at 0, 15, and 30 minutes and 1, 2, 3, 6, 14, 24, 48, 72, and 96, hours, and used to prepare total RNA. 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 |
Phoma medicaginis fungal RNA |
| Additional comments |
|
| Library developer |
Nancy L. Paiva, Samuel Roberts Noble Foundation |
| Publication reference |
|