| Title |
ESTs from pooled Medicago truncatula germinating seed |
| Library name |
NF-GS, Germinating seed |
| Library type |
EST, 5'-3' directional cDNA |
| Organism |
Medicago truncatula |
| Cultivar |
Jemalong (A17) |
| Tissue type |
Seeds |
| Developmental stage |
0, 1, 2 and 3 days after imbibing |
| 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 |
M. truncatula seeds were acid treated and placed on wet filter papers in petri dishes. Seeds were harvested at 0, 1, 2 and 3 days after acid treatment. cDNA was prepared from polyA+ enriched, pooled samples of equivalent amounts of total RNA from each time point. 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 |
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