Site Search by Google

Resources

Libraries
Human "32k" BAC Re-Array
GENSAT Collection
Vectors
Filters
Filter Interpreter Application
Mapped Clones
Non-Recombinants


Procedures
Online Mapping Resources
Download Page
Questions and Answers

Research

BAC Library Construction
ALS
Publications

 
 Home > Resources > Libraries
  CHORI-224 Honey bee BAC Library   
  Background information

The CHORI-224 library has been constructed by Chung-Li Shu and Kazutoyo Osoegawa in Pieter de Jong's laboratory, BACPAC Resources, Children's Hospital Oakland Research Institute. Agarose-embedded genomic DNA was prepared from Apis mellifera DH4 strain by Dr. Stephen Richards (Human Genome Sequencing Center Baylor College of Medicine). To create the BAC clones, genomic DNA was partially digested with a combination of EcoRI and EcoRI Methylase. The restriction fragments were size-selected using pulsed-field gel electrophoresis and then ligated into the pTARBAC2.1 vector between the EcoRI sites. The ligation products were transformed into DH10B (T1 phage-resistant) electro-competent cells (Invitrogen). The library has been arrayed into 384-well microtiter dishes and has also been gridded onto a 22x22cm nylon high-density filter for screening by probe hybridization. Each hybridization membrane represents about 18,000 distinct BAC clones, each represented in duplicate. Library construction was supported by a sub-contract from a grant awarded to Dr. J. Spencer Johnston (Department of Entomology, Texas A&M University). The collaborative effort has been organized by Daniel Weaver (President, Bee Weaver Apiaries, Inc.).


Provisional data for CHORI-224 Honey bee BAC Library:

 

Cloning Vector DNA Restriction enzyme Plate Number Total Plates Empty Wells Empty Wells (%)
pTARBAC2.1 DH4 MboI 1-96 96 355 1.0

Non-insert clones (%) Non-insert clones Non-Recombinant Clones (pUC) Recombinant clones Average Insert Size Redundancy (Genome Size: 280 Mb)
0.7 Approx. 255 N/A Approx. 36,250 158 kbp 20.5

Data on the CHORI-224 clone average insert size has been determined by Pulsed Field Gel Electrophoresis. Clone size distribution has been plotted graphically. While analyzing clones using pulse-field electrophoresis to determine the average insert size, non-insert clones containing a small deleted vector fragment consistent with sucrose resistance were observed. Further in depth characterization of the library is on going in our lab and data will be updated on our web page periodically.

Please direct questions concerning this library to either Pieter J. de Jong or Kazutoyo Osoegawa.

  Ordering & Pricing information

The library is available in several formats. Individual clones, and high-density hybridization filters  are obtainable. For ordering and shipping details, please view the ordering and pricing information  page.

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).

Note: We are aware that about 30% of unique probes from honeybee fail to find corresponding sequences in the BAC library.  This relates to the genomic composition of the honeybee genome. Please be aware of the risk that any positive clones may not be found even the library is screened with high-density filters using probes with good technical skills. It is user's responsibity to make a judgement whether or not the library should be screened. We will not be able to make any refund even the user fail to find any positive clones.

  Reference

Reference: The Honeybee Genome Sequencing Consortium (2006) Insights into social insects from the genome of the honeybee Apis mellifera. Nature 443: 931-49.

 

 

For questions related to the site, please contact webmaster.
The use of this website is subject to the terms of use.