RPCI - 24 Mouse (C57BL/6J Male) BAC Library

The RPCI-24 BAC library has been constructed in our laboratory from tissues derived from single male C57BL/6J mouse. Spleen and brain genomic DNA samples were isolated and partially digested with MboI. Size-selected MboI fragments were cloned into the pTARBAC1 vector between the BamHI sites. The ligation products were transformed into DH10B electrocompetent cells (BRL Life Technologies). The library has been arrayed into 384-well microtiter dishes and also gridded onto 22x22cm nylon high-density filters for screening by probe hybridization. Each hybridization membrane represents over 18,000 distinct mouse BAC clones, stamped in duplicate.

The RPCI - 24 Male (C57BL/6J) Mouse BAC Library

Segments	Cloning Vector	DNA	Restriction enzyme	Plate Numbers	Total Plates	Empty Wells	Empty Wells (%)
1	pTARBAC1	Spleen	MboI	1-161, 168-177	288	3464	3.1
		Brain	MboI	162-167, 178-288
2	pTARBAC1	Brain	MboI	289-313	288	3951	3.6
		Spleen	MboI	314-576
Total Library					576	7415	3.35

Segments	Non-insert Clones (%)	Non-insert clones	Non-Recombinant Clones (pUC)	Recombinant Clones	Average Insert Size	Redundancy (Genome size: 3,000 Mb)
1	4.3	4607	1	102,521	160 Kbp	5.5
2	0.5	533	0	106,108	150 Kbp	5.3
Total Library	2.4	5140	1	208,629	155 Kbp	10.8

click here for a legend of the previous tables.

Data on the RPCI-24 clone average insert size (Segment 1, Segment 2) has been determined by Pulsed Field Gel Electrophoresis. Clone size distribution has been plotted graphically.