|common Name:||P. euphratica BIBAC|
|Genome Size (Mb):||495.87|
|Insert Size (Kb):||127|
We have constructed a BIBAC library for P. euphratica as a resource to share with every academic researchers. This BIBAC library is stored at –80°C in State Key Laboratory of Tree Genetics and Breeding, Chinese Academy of Forestry. The information of this BIBAC library is provided as follows:
Cotyledons (20 g) were ground in liquid nitrogen and high molecular weight DNA was isolated from nuclei prepared as described previously1. DNA (1 mg) was partially digested with 2 U of BamHI (New England Biolabs, USA) for 3 h at 37°C in a reaction volume of 40 µL, and restriction fragments of 100–300 kb were selected by pulsed-field gel electrophoresis and cloned into the BIBAC vector pCLD04541 (kindly provided by Dr. C. Dean, John Innes Institute, Norwich, UK) as previously described2.
The recombinant DNAs were transformed into Escherichia coli DH10B competent cells (Invitrogen, USA) with the Cell Porator and Voltage Booster electroporation system (Life Technologies, USA) and transformed cells were selected on Luria-Bertani (LB) medium containing 15 µg mL–1 tetracycline2. The BIBAC library constructed for P. euphratica comprised 55,296 clones arrayed in 144 × 384 microplates containing 50 µL of LB medium and stored at –80°C. To estimate the size of the inserts in the library, insert DNAs were isolated from 114 randomly selected clones and resolved by pulsed-field gel electrophoresis following digestion with NotI. The insert size ranged from 50 to 150 kb, we estimated that the library had a mean insert size of 127 kb. The genome size of P. euphratica was estimated to be approximately 495.87 Mb3, thus this BIBAC library represented the equivalent of approximately 14× the P. euphratica genome.
Database for P. euphratica BIBAC clones is under construction and could be queried soon. We are willing to share BIBAC clones for free with academic researchers for non-commercial purposes. It’s important to note that you need to pay the freight, and we can propagate them for free if less than 5 clones are requested, otherwise we just provide clones and you need to propagate them by yourself. Please feel free to contact Zhou Jing (email: email@example.com) if you have any question about this BIBAC library.
1. Zhang, H. B., Zhao, X., Ding, X., Paterson, A. and Wing, R. 1995, Preparation of megabase-size DNA from plant nuclei. Plant Journal 7, 175-184.
2. Zhang, H. B., Scheuring, C. F., Zhang, M., et al. 2012, Construction of BIBAC and BAC libraries from a variety of organisms for advanced genomics research. Nature Protocols, 7, 479-499.
3. Ma, T., Wang, J., Zhou, G., et al. 2013, Genomic insights into salt adaptation in a desert poplar. Nature communications, 4, 2797.