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 Home > Resources > Libraries
   CHORI-251: Chimpanzee (M) (Pan troglodytes) BAC Library   

  Background information

The CHORI-251 Chimpanzee BAC library has been constructed at the Children's Hospital Oakland Research Institute, BACPAC Resources, by Baoli Zhu for segments 1 and 2, Michael Nefedov for segments 3 and 4 using the cloning techniques developed in our laboratory (Osoegawa et al., 1998). DNA was isolated from white blood cells obtained from a male chimpanzee (Pan troglodytes, "Clint", Yerkes #C0471; birthdate: 6-6-80). Agarose embedded high molecular weight DNA was partially digested with a combination of EcoRI restriction enzyme and EcoRI Methylase for segments 1 and 2, and with MboI for segments 3 and 4. Size fractionated DNA was cloned into the pTARBAC2.1 vector between the EcoRI sites or pTARBAC1.3 vector between the BamHI sites. The ligation products were transformed into DH10B (T1 resistant) electrocompetent cells (BRL Life Technologies). The library has been arrayed into 384-well microtiter dishes and also gridded onto 24 22x22cm nylon high-density filter for screening by probe hybridization. Each hybridization membrane represents over 18,000 distinct Chimpanzee BAC clones, stamped in duplicate. BAC library construction was funded by a grant from NIH (#HG01165-06).

Segment

1

2

3

4

All

Vector

pTARBAC2.1

pTARBAC2.1

pTARBAC1.3

pTARBAC1.3

Restriction Enzyme

EcoRI/EcoRI Methylase

EcoRI/EcoRI Methylase

MboI

MboI

N/A

DNA Source

blood

blood

blood

blood

blood

Plate Numbers

1-288

289-576

577-864

865-1152

1-1152

Plate Count

288

288

288

288

1152

Empty Wells

1630 (1.47%)

1399 (1.27%)

2295 (2.08%)

2251 (2.04%)

7595 (1.72%)

Non-Recombinant Clones

552 (0.51%)

653 (0.6%)

N/A

N/A

1205 (0.28%)

Non-Insert Clones

Approx. 3923 (3.6%)

Approx. 6552 (6%)

Approx. 2762 (7/275, 2.55%)

Approx. 3954 (10/274, 3.65%)

Approx. 20000 (4.6%)

Recombinant Clones

104487

101988

105535

104387

413568

Average Insert Size

164 Kbp

176 Kbp

190 Kbp

180 Kbp

178 Kbp

Genomic Coverage

5.2X

5.4X

6.3X

6X

22.9X



click here for a legend of the previous tables.

No clones were found to be non-recombinant after analysis of CHORI-251 using overgo probes specific for the puc19 fragment.

Data on the CHORI-251 clone insert size distribution has been determined by pulsed-field gel Electrophoresis. Clone Size Distribution has been plotted graphically.

The library is available as library plates, individual clones, and high-density hybridization sets (see Filter Availability)

Individual clones identified after screening can be obtained as stab cultures at a cost of $80 per clone in addition to a $35 set-up charge per shipment. Shipping will be done by Federal Express and charged to the recipients account number.

With respect to the scheduling of shipments of arrayed library copies, we would like to make you aware of our policy for array distribution.
Please visit the ordering information page if you want to place an order.

Academic and commercial users interested in a copy of the BAC library should contact Pieter J. de Jong (pdejong@chori.org, fax: (510) 450-7924). For hybridization membranes, please contact BACPAC Resources (BACPACorders@chori.org).

 

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