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 Home > Resources > Libraries
  CHORI-257 Grey Mouse Lemur
(Microcebus murinus) (Female) BAC Library
  


The CHORI-257 mouse lemur BAC library has been constructed by Michael Nefedov in Pieter de Jong’s laboratory at BACPAC Resources, Children’s Hospital Oakland Research Institute. The preparation of the library followed the cloning approach developed in our laboratory (Osoegawa et al., 1998). Kidney was obtained from an adult female mouse lemur (Microcebus murinus) through John Allman, Division of Biology 216-76, Caltech, Pasadena, CA 91125. The local identification number for this animal is #8. A skin biopsy and a blood sample have been sent to the Coriell Cell Repositories to establish cell lines as part of the Integrated Primate Biomaterials and Information Resource (IPBIR) initiative. High-molecular-weight DNA was isolated from a kidney cell suspension prepared in a Dounce Homogenizer and the cells were embedded in agarose to stabilize the DNA. The embedded DNA was partially digested with a combination of EcoRI restriction enzyme and EcoRI methylase and size fractionated by pulsed-field electrophoresis. 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 576 “384-well” microtiter dishes and also gridded onto twelve 22x22cm nylon high-density filters for screening by probe hybridization. Each hybridization membrane represents about 18,000 distinct mouse lemur BAC clones, stamped in duplicate. Library characterization was performed by Qing Cao, Teresa Ren and Kazutoyo Osoegawa. This work was funded by NIH grant HG025323-01.

Segment 1 2 Total
Cloning Vector pTARBAC2.1 pTARBAC2.1  
DNA Source Kidney Cell Kidney Cell  
Restriction Enzyme EcoRI EcoRI  
Plate Range 1-288 289-576 1-576
Total 384-Well Dishes 288 288 576
Empty Wells (%) 1,000 (0.9%) 2,400 (2.2%) 3,500 (1.5%)
Non-insert Clones (%) 4/287 (1.4%) 7/274 (2.6%) 11/561 (2.0%)
Vector Clones (%) 0/110K (0) 0/110K (0) 0/220K (0)
Recombinant Clones ~108K ~105K ~213K (>96% of wells)
Insert size (Average) ~210Kbp ~220Kbp ~214Kbp
Standard Deviation 57Kbp 55Kbp 56Kbp
Library Redundancy ~7.6 fold ~7.7 fold ~15 fold


Click here for a legend of the previous tables.
Data on the CHORI-257 clone average insert size has been determined by Pulsed Field Gel Electrophoresis. Clone size distribution has been plotted graphically.
Hybridization filters can be ordered per complete set of 12 membranes (price per set is 12 times $200.00) or per segment (6 times $200.00). Shipping will be done by Federal Express and charged to recipient's 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).

Please contact Pieter J. de Jong or Kazutoyo Osoegawa for questions related to this library.

 

 

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