| Title |
ESTs from Medicago truncatula roots |
| Library name |
NF-RT, Developing root |
| Library type |
EST, 5'-3' directional cDNA |
| Organism |
Medicago truncatula |
| Cultivar |
Jemalong (A17) |
| Tissue type |
Root |
| Developmental stage |
Pooled developmental |
| 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 |
Total RNA was extracted from non-nodulated roots of plants grown in Perlite saturated with a complete fertilizer containing 1mM nitrate. Samples were taken at four time points (approximately two days, one, two and six weeks post germination) representing early seedling growth to nitrogen limitation. 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. |
| ' 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 |
Nancy L. Paiva, Samuel Roberts Noble Foundation |
| Publication reference |
|