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CHORI-277: Female Gorilla "Kamilah" (Gorilla gorilla gorilla) BAC Library |
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The CHORI-277 Gorilla BAC Library has been generated by Baoli Zhu, in Pieter de Jong’s laboratory at BACPAC Resources, Children’s Hospital Oakland Research Institute. The library preparation followed the general cloning approach developed in our laboratory Osoegawa et al., 1998. Blood from female gorilla “Kamilah” was kindly provided by Dr. Oliver Ryder from the Zoological Society of San Diego's center for Conservation and Research for Endangered Species (CRES). White blood cells were embedded in 0.5% InCert agarose and proteins were removed by a detergent/proteinaseK treatment. The resulting agarose-embedded high molecular weight DNA was partially digested with a combination of EcoRI restriction enzyme and EcoRI methylase then size fractionated by pulsed-field gel electrophoresis. DNA fragments from the appropriate size fraction were cloned into the pBACgmr 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 576, 384-well microtiter dishes and has subsequently been gridded onto 12, 22x22cm nylon high-density filters for screening by probe hybridization. Each hybridization membrane represents over 18,000 distinct BAC clones, stamped in duplicate.
The preparation of this library was facilitated through a subcontract from the Sanger Institute, funded by The Wellcome Trust. The BAC library will be used as a key reagent for gorilla genome project. BAC-derived data has been co-assembled with whole-genome shotgun sequence data derived from small insert and fosmid libraries (fosmids: CHORI-1277), each derived from the same gorilla "Kamilah". The BAC library is in the public domain and clones can be obtained following the "Ordering Instructions" on our home page. About 354,000 BAC-end-sequences can be found in the NCBI Trace Archive using the following query string: [species_code="GORILLA GORILLA" and center_name="WUGSC" and strategy="CLONEEND" and library_ID="CH277"]. Data for CHORI-277 Gorilla BAC Library:
Segment | 1 | 2 | All | Vector | pBAC-GMR | pBAC-GMR | pBAC-GMR | Restriction Enzyme | EcoRI/EcoRI Methylase | EcoRI/EcoRI Methylase | EcoRI/EcoRI Methylase | DNA Source | blood | blood | blood | Plate Numbers | 1-288 | 289-576 | 1-576 | Plate Count | 288 | 288 | 576 | Empty Wells | 2958 (2.67%) | 2928 (2.65%) | 6084 (2.75%) | Non-Recombinant Clones | N/A | N/A | N/A | Non-Insert Clones | Approx. 2002 (1.86%) | N/A | N/A | Recombinant Clones | 105632 | 107664 | 215100 | Average Insert Size | 163 Kbp | 154 Kbp | 159 Kbp | 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-277 using overgo probes specific for the puc19 fragment.
Data on the CHORI-277 clone insert size distribution has been determined by pulsed-field gel Electrophoresis. Clone Size Distribution has been plotted graphically.
The BAC library has been gridded onto 22x22cm positively charged nylon filters for hybridization screening purposes. Each filter contains 36,864 colonies which represents 18,432 independent clones spotted in duplicate in a 4x4 clone array. Clones can be identified through screening by purchasing "high-density colony" hybridization filters or by utilizing our "fee for service" "Library Screening Services" . For questions about clones and hybridization membranes, please contact BACPAC Resources ([email protected]).
Please address questions concerning this library to Pieter J. de Jong.
Ordering & Pricing information
The complete copy of this BAC library is not available. The library is available in several formats. Individual clones, and high-density colony filters for hybridization screening experiments hybridization filter. For ordering and shipping details, please click here. Please address questions concerning this library to Pieter J. de Jong.
Reference
Scally A, Dutheil JY, Hillier LW, Jordan GE, Goodhead I, et al. (2012) Insights into hominid evolution from the gorilla genome sequence. Nature 483: 169-175.
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