Mouse Genomic BAC library - RPCI-23 (RP23)
The RPCI-23 (RP23) BAC Library was constructed in Pieter de Jong's lab (Children's Hospital Oakland Research Institute) by visiting scientists Kazutoyo Osoegawa and Minako Tateno (RIKEN Tsukuba Life Science Center). From pooled tissues derived from three female C57BL/6J mice, kidney and brain genomic DNA samples were isolated and partially digested with a combination of EcoRI and EcoRI Methylase. Size-selected EcoRI fragments were cloned into the vector pBACe3.6 (pBACe3.6 sequence) at the EcoRI sites. The ligation products were transformed into DH10B electro-competent cells (BRL Life Technologies).
The library was arrayed into 384-well microtitre dishes and also gridded onto 10 22x22cm nylon high-density filters for screening by probe hybridization
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Library: RPCI-23 (RP23) DNA source: C57BL/6L kidney & brain Vector: pBACe3.6 |
|
Segment | Cloning
| Total
clones | Number
of plates | Non-insert
clones(%) | Insert
size
(kbp's) | Genomic
representation |
| 1 | EcoRI | 80,661 | 240 | 11.3 | 198 | 5.3x |
| 2 | EcoRI | 89,973 | 240 | 1.3 | 197 | 5.9x |
| Total | - | 170,634 | 480 | 6 | 197 | 11.2x |
For more information on how the library was constructed please consult:
Osoegawa K, Tateno M, Woon PY, Frengen E, Mammoser AG, Catanese JJ, Hayashizaki Y, de Jong PJ.Bacterial artificial chromosome libraries for mouse sequencing and functional analysis. Genome Res. 2000 Jan;10(1):116-28.
Or visit the website: http://bacpac.chori.org/
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