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 Home > Resources > Libraries
   CHORI-264: White-throated Sparrow (Zonotrichia albicollis) BAC Library   

Background information

The CHORI-264, White-throated Sparrow, BAC Library was generated by Boudewijn ten Hallers 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). Frozen kidney tissue obtained from a female sparrow, sample ID:822, was kindly provided by James W. Thomas, PhD in the Department of Human Genetics at Emory University School of Medicine, 615 Michael St., Suite 301, Atlanta, GA 30322. The frozen kidney tissue was ground to a fine powder, resuspended in chromatin isolation buffer then embedding in 0.5% InCert agarose. The proteins were removed by a detergent/proteinaseK treatment and the resulting high molecular weight DNA 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

kidney/brain

kidney/brain

kidney/brain

Plate Numbers

1-288

289-528

1-528

Plate Count

288

240

528

Empty Wells

3106 (2.81%)

2464 (2.67%)

5570 (2.75%)

Non-Recombinant Clones

N/A

N/A

N/A

Non-Insert Clones

Approx. 430 (1/252, 0.4%)

Approx. 395 (1/227, 0.44%)

Approx. 828 (2/479, 0.42%)

Recombinant Clones

107056

89301

196354

Average Insert Size

146 Kbp

141 Kbp

144 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-264 using overgo probes specific for the puc19 fragment.

Data on the CHORI-264 clone insert size distribution has been determined by pulsed-field gel Electrophoresis. Clone Size Distribution has been plotted graphically.




Questions related to this library should be sent to Pieter J. de Jong (pdejong@chori.org, fax: (510) 450-7924).

Posted: September 19, 2008
Revised: October 22, 2008

 

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