LBNL-1 Rabbit BAC Library

Name	# of plates	% empty-well	% no-insert	# of recombinants	Avg. insert size	Coverage
LBNL-1	322	1%	1.3%	120,804	175 kb	7.0X

This BAC library was constructed by Ze Peng in the laboratory of Jan-Fang Cheng at Lawrence Berkeley National Laboratory using the genomic DNA isolated from white blood cells of a New Zealand White rabbit (Oryctolagus cuniculus). The rabbit DNA was partially digested with EcoRI and EcoRI methylase, size selected, and cloned into the pBACe3.6 vector according to the protocol described in Osoegawa et al. (1998). The ligated DNA was then transformed into DH10B electro-competent cells (Invitrogen 18290-015). This library so far contains 322 384-well plates with an average insert size of 175 Kb. The construction of this library was supported by a grant (HL66728-01) from the National Heart, Lung and Blood Institute of NIH. This library is characterized by the following criteria:

(1) Size distribution:

(2) BAC end sequencing (Collaboration with Dr. Shaying Zhao of TIGR):

Total sequencing attempts: 863
Good sequences: 700
Avg reads: 463bp
Avg Q20 bases: 420bp
Success sequences: 81%
Low quality sequences: 16%
Vector sequences: 11 (1.3%)
E coli sequences: 0
Background: 9 (1%)

(3) Library screening:
We have hybridized the library filters with 27 probes representing 18 genomic intervals to estimate the randomness of the coverage. All probes have identified positives ranging from 1 to 12 hits per probe. The average number of hits is 5.