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CHORI-560: H1 Human Embryonic BAC Library |
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The CHORI-560 H1 Human Embryonic Stem cell line BAC Library was constructed from a suspension of H1 embryonic stem cells. The library was generated by Yongli Bai and Maxim Koriabine in Pieter de Jong’s laboratory at BACPAC Resources, Children’s Hospital Oakland Research Institute. Procedures followed the cloning approach developed in our laboratory (Osoegawa et al., 1998). The cells were provided by the Baylor College of Medicine. The cell suspension was embedding in 0.5% InCert agarose and proteins were removed by a detergent/proteinaseK treatment. The resulting high molecular weight DNA was partially digested using a combination of EcoRI restriction enzyme and EcoRI methylase enzyme then size fractionated by pulsed-field gel electrophoresis. DNA fragments from the appropriate size fraction were cloned into the pSG1473 vector between the two EcoRI sites. The ligation products were transformed into MDS42recA trfA Blue electro-competent cells (Scarab Genomics). The library has been arrayed into 720 (384-well) microtiter dishes.Segment | 1 | 2 | All | Vector | pSG1473 | pSG1473 | pSG1473 | Restriction Enzyme | EcoRI/EcoRI Methylase | EcoRI/EcoRI Methylase | EcoRI/EcoRI Methylase | DNA Source | cell line | cell line | cell line | Plate Numbers | 1-336 | 337-720 | 1-720 | Plate Count | 336 | 384 | 720 | Empty Wells | N/A | N/A | 558 (0.2%) | Non-Recombinant Clones | N/A | N/A | N/A | Non-Insert Clones | Approx. 361 (0.28%) | Approx. 840 (0.57%) | Approx. 1159 (3/712, 0.42%) | Recombinant Clones | 128663 | 146616 | 274763 | Average Insert Size | 147 Kbp | 146 Kbp | 146 Kbp | Genomic Coverage | N/A | N/A | N/A |
Data on the CHORI-560 clone insert size distribution has been determined by pulsed-field gel Electrophoresis. Clone Size Distribution has been plotted graphically.
No clones were found to be non-recombinant after analysis of CHORI-560 using overgo probes specific for the puc19 fragment.
Question related to this library should be sent to Pieter J. de Jong ([email protected], fax: (510) 450-7924).
Posted: October 17, 2007
Revised: April 11, 2008
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