RPCI - 21 Female (129S6/SvEvTac) Mouse PAC Library
The RPCI-21 PAC Library has been constructed in our laboratory by Kazutoyo Osoegawa. PAC library construction was funded by grants from the US Department of Energy (#DE-FG02-94ER61883) and the National Human Genome Research Institute (NHGRI, NIH) (#1R01RG01165-03). female 129S6/SvEvTac mouse spleen genomic DNA (partially MboI digested) was cloned between the Bam HI sites of the pPAC4 vector. The ligation products were transformed into DH10B electrocompetent cells (BRL Life technologies). The library has been arrayed into 384 well microtiter dishes and then gridded onto 22x22cm nylon high density hybridization filters for screening by probe hybridization. Each hybridization membrane represents over 18,000 distinct mouse PAC clones, stamped in duplicate.
The Total RPCI-21 Mouse PAC Library:
Clones Picked |
Empty Wells |
% Empty |
Non Recombinant Clones |
Non Recombinant Clones (%) |
Total Clones |
Average Insert Size |
384 Well Plate Numbers |
| 258048 | 6565 | 2.5 | 1312 | 0.5 | 250040 | 137 Kbp | 1-672 |
Segments |
Clones Picked |
Empty Wells |
Non Recombinant (Total) |
Non Recombinant Clones (%) |
% Empty |
Total Clones |
Average Insert Size |
384 Well Plate Numbers |
Gemonic Coverage |
1 |
129836 | 3868 | 650 | 0.5 | 3.0 | 125318 | 127 Kbp | 1-336 | 5.3X |
2 |
132473 | 2912 | 662 | 0.5 | 2.2 | 128899 | 147 Kbp | 337-672 | 6.3X |
click here for a legend of the previous tables.
The average insert size data has been determined
by Pulsed field Gel Electrophoresis.
Reference:
Osoegawa, K. et al. (2000) Bacterial Artificial Chromosome Libraries for Mouse
Sequencing and Functional Analysis. Genome Research 10: 116-128
Reprints availiable upon request.