Legume BAC library information Resources Page https://legumeinfo.org/data/v2/Cajanus/cajan/supplements/ICPL87119.bac1.GpKx/ Pigeonpea (Cajanus cajan) CAJANUS CAJAN LIBRARY CccABb Library name: CccABb Plant genotype: cultivar "Asha" Vector: Epicenter Copy Control pCC1BAC Host strain: Transformax EPI30 Enzyme: BamHI Average insert size: 115 kbp Number of clones: 34,560 Estimated genome coverage: 5.2X Contact: Doug Cook Description: Pigeonpea (Cajanus cajan) accession Asha was grown under greenhouse conditions to the seedling stage and transferred to continuous darkness for 2 days prior to use. Nuclei were isolated and embedded in low melting point agarose, restriction digested with BamHI and size selected by means of two rounds of pulsed field gel electrophoresis. Large size DNA fragments were ligated in vector pCC1BAC and transformed by electroporation in to Epicenter's E. coli EPI300-T1R cells. 34,560 clones from were obtained, with an average insert size of 115 kbp. BAC end sequences were obtained from ~25,000 clones. To obtain clones from this library, please contact Doug Cook at UC Davis Email: drcook@ucdavis.edu Phone: 530-754-6561 Pigeonpea (Cajanus cajan) CAJANUS CAJAN LIBRARY CccABa Library name: CccABa Plant genotype: cultivar "Asha" Vector: Epicenter Copy Control pCC1BAC Host strain: Transformax EPI30 Enzyme: HindIII Average insert size: 120 kbp Number of clones: 34,560 Estimated genome coverage: 5.5X Contact: Doug Cook Description: Pigeonpea (Cajanus cajan) accession Asha was grown under greenhouse conditions to the seedling stage and transferred to continuous darkness for 2 days prior to use. Nuclei were isolated and embedded in low melting point agarose, restriction digested with Hind III and size selected by means of two rounds of pulsed field gel electrophoresis. Large size DNA fragments were ligated in vector pCC1BAC and transformed by electroporation in to Epicenter's E. coli EPI300-T1R cells. 34,560 clones from were obtained, with an average insert size of 120 kbp. BAC end sequences were obtained from ~25,000 clones. To obtain clones from this library, please contact Doug Cook at UC Davis Email: drcook@ucdavis.edu Phone: 530-754-6561 Chickpea (Cicer arietinum) CICER ARIETINUM LIBRARY CAA1Ba Library name: CAA1Ba Plant genotype: ICC4958 Vector: Epicenter Copy Control pCC1BAC Host strain: Transformax EPI30 Enzyme: HindIII Average insert size: 100 to 130 kbp Number of clones: 55,680 Estimated genome coverage: 8.5X Contact: Doug Cook Description: Chickpea (Cicer arietinum) accession ICC4958 was grown under greenhouse conditions for 6 weeks and transferred to continuous darkness for 2 days prior to use. Nuclei were isolated and embedded in low melting point agarose, restriction digested with Hind III and size selected by means of two rounds of pulsed field gel electrophoresis. Large size DNA fragments were ligated in vector pCCBAC1H and transformed by electroporation in to Epicenter's E. coli EPI300-T1R cells. 55,680 clones from were obtained, with most inserts ranging from 100 to 130 kbp. BAC end sequences were obtained from ~25,000 clones. To obtain clones from this library, please contact Doug Cook at UC Davis Email: drcook@ucdavis.edu Phone: 530-754-6561 Cowpea (Vigna unguiculata) VIGNA UNGUICULATA LIBRARY VUH2 Library name: VUUBBa (VUH2) Plant genotype: Blackeye 5 line 9405C Vector: Epicenter Copy Control pCC1BAC Host strain: Transformax EPI30 Enzyme: HindIII Average insert size: 80-110 kbp Number of clones: 36,864 Estimated genome coverage: 5X Contact: Doug Cook Description: Cowpea (Vigna unguiculata) cultivar Blackeye 5 was crossed to cowpea line Arlington, the source of the dominant Cpa gene for resistance to Cowpea mosaic virus. Blackeye 5 was the recurrent parent in an eight-fold backcross initiated with the Blackeye 5 x Arlington cross. 9405C is a line (genotype Cpa/Cpa) derived from several generations of selfing from a single seed derived from the backcross series. 9405C seeds were germinated in sterile soil under greenhouse conditions for 3-4 days. As the seedling hook was about to break ground, the pots were covered to exclude almost all light for approximately 36 hours. The yellow-green primary leaves were harvested and snap-frozen under liquid nitrogen. Nuclei were isolated in a high pH Tris buffer to counteract tissue acidity. The nuclei were embedded in low melting point agarose, digested with proteinase K, restriction digested with Hind III and size selected by means of two rounds of pulsed field gel electrophoresis. Large size DNA fragments were ligated into HindIII-cut vector pCC1BACH and transformed by electroporation in to E. coli Transformax EPI300 (Epicentre) cells. 130 clones were assessed for insert size by excision of the insert by NotI digestion and CHEF gel analysis of the products. Most inserts were 80-110 kbp with an average insert size of 100 kbp. 36,864 clones were collected and BAC end sequences were obtained for ~27,000 clones. Cowpea (Vigna unguiculata) VIGNA UNGUICULATA LIBRARY VUH1 Library name: VUUBBa (VUH1) Plant genotype: Blackeye 5 Vector: Epicenter Copy Control pCC1BAC Host strain: Transformax EPI30 Enzyme: HindIII Average insert size: 65 kbp Number of clones: 73,728 Estimated genome coverage: 8X Contact: Doug Cook Description: Seed of cowpea (Vigna unguiculata) cultivar Blackeye 5 was germinated in sterile soil under greenhouse conditions for 3-4 days. As the seedling hook was about to break ground, the pots were covered to exclude almost all light for approximately 36 hours. The yellow-green primary leaves were harvested and snap-frozen under liquid nitrogen. Nuclei were isolated in a high pH Tris buffer to counteract tissue acidity. The nuclei were embedded in low melting point agarose, digested with proteinase K, restriction digested with Hind III and size selected by means of two rounds of pulsed field gel electrophoresis. Large size DNA fragments were ligated into HindIII-cut vector pCC1BACH and transformed by electroporation in to E. coli Transformax EPI300 (Epicentre) cells. Approximately 50 clones were assessed for insert size by excision of the insert by NotI digestion and CHEF gel analysis of the products. Most inserts were 40-90 kbp with an average insert size of 65 kbp. 73,728 clones were collected and BAC end sequences were obtained for ~2300 clones. To obtain clones from this library, please contact Doug Cook at UC Davis Email: drcook@ucdavis.edu Phone: 530-754-6561 Peanut (Arachis hypogaea) ARACHIS HYPOGAEA LIBRARY AHT1 Library name: AHT1 Plant genotype: cultivar "Tifrunner" Vector: pSMART-BAC Host strain: E. coli DH10B Enzyme: Sheared DNA Average insert size: 100 kbp Number of clones: 110,000 Estimated genome coverage: ~4X Contact: Doug Cook Description. Peanut (Arachis hypogaea) cultivar Tifrunner was grown under greenhouse conditions for 6 weeks and transferred to continuous darkness for 2 days prior to use. High molecular weight DNA was isolated and random sheared DNA was size selected based on proprietary methods from Lucigen Corp. Large size DNA fragments were ligated in vector pSMART-BAC and transformed by electroporation in to Epicenter's E. coli 10G (DH10B) BAC-Optimized cells. Approximately 110,000 clones were obtained, with most inserts averaging 100 kbp. To obtain clones from this library, please contact Doug Cook at UC Davis Email: drcook@ucdavis.edu Phone: 530-754-6561 Red Bud (Cercis chinensis) CERCIS CHINENSIS LIBRARY CC1 Library name: CC1 Plant genotype: Cercis chinensis from the US National Arboretum collection Vector: Epicenter Copy Control pCC1BAC Host strain: Transformax EPI30 Enzyme: HindIII Average insert size: 100 kbp Number of clones: 34,560 Estimated genome coverage: 10X Contact: Doug Cook Description: Red Bud (Cercis chinensis) tissue was collected as newly emerged leaves from a single individual tree at the US National Arboretum collection. Leaves were harvested in the spring and frozen directly in liquid nitrogen. Nuclei were isolated and embedded in low melting point agarose, restriction digested with HindHI and size selected by means of two rounds of pulsed field gel electrophoresis. Large size DNA fragments were ligated in vector pCC1BAC and transformed by electroporation in to Epicenter's E. coli EPI300-T1R cells. 34,560 clones from were obtained, with an average insert size of 100 kbp. To obtain clones from this library, please contact Doug Cook at UC Davis Email: drcook@ucdavis.edu Phone: 530-754-6561