Legume bac library information Resources Page




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Legume BAC library information Resources Page
http://www.comparative-legumes.org/pages/resources

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


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