Barley Embryo Sack Expression cDNA
cDNA for this library was synthesised from barley embryo sack sampled 0-10 days after flowering (Cultivar: Barke). The main library of 21500 clones was re-arrayed into the sub-library BES1824 containing 4100 putative expression clones. Average insert size is 1 kb. Library generation and sequencing was granted in context of GABI; data are also accessible at http://www.gabipd.org/
About 3,900 cDNA clones are available in the E. coli expression vector pQE30NST. The vector is suitable for expression of N-terminal His-tagged proteins.
As a source of recombinant barley proteins.
Note: Due to a cloning artefact, in most cases the SalI site is NOT present, as well as the SalI Adapter used for cloning. To excise the insert, restriction sites upstream SalI should be used (e.g. BamHI).
||Barley Embryo Sack Expression cDNA (1)
||Species: Hordeum vulgare; Tissue: embryosack (endosperm with embryo)
||Cloning Site 3s: SalI; Cloning Site 5s: NotI
||Host Species: E. coli; Host Strain: SCS1 (Stratagene)
||Growth Conditions medium: 2YT; Growth Conditions antibiotic: Amp (100 µg/ml); Growth Conditions antibiotic: Kan (15 µg/ml)
Protocols / Clone Handling
Plates- contain LB broth with 8% glycerol and antibiotic* and are sent on dry ice. These should be stored at -70°C. Individual clones have been streaked onto LB agar containing antibiotic*. Please store them at 4°C (not in a freezer).
As soon as possible, re-streak the clones onto the same medium and incubate overnight at 37°C to obtain single colonies.
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