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CHORI-263: Budgerigar (Melopsittacus undulatus) BAC Library |
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The CHORI-263, Budgerigar, BAC Library was generated by Yongli Bai and Maxim Koriabine in Pieter de Jong’s laboratory at BACPAC Resources, Children’s Hospital & Research Center Oakland. Procedures followed the cloning approach developed in our laboratory (Osoegawa et al., 1998). Blood was obtained from a female budgerigar named Roz. The blood sample was kindly provided by Timothy F. Wright, PhD in the Department of Biology MSC 3AF, New Mexico State University, Las Cruces, NM 88003. White blood cells were embedded in 0.5% InCert agarose and proteins were removed by a detergent/proteinaseK treatment. The agarose embedded, high molecular weight DNA was partially digested using a combination of EcoRI restriction enzyme and EcoRI methylase enzyme. The DNA was size fractionated by pulsed-field gel electrophoresis and DNA fragments from the appropriate size fraction were cloned into the pTARBAC2.1 vector between the two EcoRI sites. The ligation products were transformed into DH10B (T1-resistant) electro-competent cells (Invitrogen). The library has been arrayed into 528 (384-well) microtiter dishes and has subsequently been gridded onto 11 22x22cm nylon high-density filters for screening by probe hybridization. Each hybridization membrane represents over 18,000 distinct BAC clones, stamped in duplicate.
Segment | 1 | 2 | All | Vector | pTARBAC2.1 | pTARBAC2.1 | pTARBAC2.1 | Restriction Enzyme | EcoRI/EcoRI Methylase | EcoRI/EcoRI Methylase | EcoRI/EcoRI Methylase | DNA Source | blood | blood | blood | Plate Numbers | 1-288 | 289-528 | 1-528 | Plate Count | 288 | 240 | 528 | Empty Wells | 2373 (2.15%) | 2987 (3.24%) | 5360 (2.64%) | Non-Recombinant Clones | N/A | N/A | N/A | Non-Insert Clones | N/A | N/A | N/A | Recombinant Clones | 108219 | 89173 | 197392 | Average Insert Size | N/A | N/A | N/A | Genomic Coverage | N/A | N/A | N/A |
Click here for legend of the previous table.
No clones were found to be non-recombinant after analysis of CHORI-263 using overgo probes specific for the puc19 fragment.
Data on the CHORI-263 clone insert size distribution is currently not available for this library and will be posted as soon as available.
Questions related to this library should be sent to Pieter J. de Jong ([email protected], fax: (510) 450-7924).
Posted: September 24, 2008
Revised:
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