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CopyControl™ pCC1BAC? (Hind III Cloning-Ready) Vector
CopyControl™ pCC1BAC? (Hind III Cloning-Ready) Vector
货号
CBAC311H
规格
375 ng
存储温度
-20℃
产品详情
Applications
- Preparation of BAC libraries that can be maintained at single copy number and induced to high-copy number as needed.
CopyControl™ BAC Cloning Kits* are complete kits containing all molecular biology reagents needed for constructing high-quality, large-insert Bacterial Artificial Chromosome (BAC) libraries. Each kit has enough reagents to make a complete human genome size library at 10X coverage. The kits feature the CopyControl™ pCC1BAC™ Vector which contains the E. coli F-factor single-copy origin of replication and oriV, a second origin of replication, which can be used to selectively induce to high copy number.
To maintain stability, CopyControl BAC clones are grown at very low copy number during the initial cloning and screening processes. Then, immediately before DNA purification, clones can be induced to 10-20 copies per cell within 2 hours by adding CopyControl Induction Solution to the culture, for higher yields of BAC DNA. The short, 2-hour induction of CopyControl BAC clones to high-copy number does not decrease their stability, based on analysis of Hind III restriction patterns of a large number of induced versus uninduced clones that ranged in size up to ~200 kb. Alternatively, the CopyControl BAC Autoinduction Solution can be used. Autoinduction Solution is added to the culture medium prior to inoculation, and also increases the cell growth leading to higher yield than standard Induction Solution. CopyControl BAC Cloning Kits reduce the time and labor required to construct a BAC library.
Phage T1-resistant TransforMax™ EPI300™-T1R Electrocompetent E. coli cells, or standard TransforMax™ EPI300™ Electrocompetent E. coli cells, which are required for induction of CopyControl BAC clones to 10-20 copies per cell, are available separately.
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Benefits
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Figure 3 (click to enlarge). The CopyControl™ BAC Cloning Kits significantly reduce the time and labor needed to construct a BAC library compared to standard methods. |
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Figure 1. DNA from induced (+) and uninduced (-) cultures of 4 CopyControl™ BAC clones. An equal number of cells were processed from both induced and uninduced cultures. Samples were resolved on a 0.8% agarose gel and stained with SYBR® Gold (Invitrogen). The induced cultures yielded >15-fold more CopyControl BAC clone DNA. M=BAC-Tracker™ Supercoiled DNA ladder. |
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Figure 2. Not I digestion of 2 µl of DNA from induced (+) and uninduced (-) cultures of 4 CopyControl™ BAC clones. After resolving by PFGE and staining with SYBR® Gold (Invitrogen), clone sizes were: BAC1=130 kb, BAC2=125 kb, BAC3=140 kb and BAC4=145 kb. M=Pulsed-Field Gel Marker. |
*Covered by issued and/or pending patents.
SYBR® is a registered trademark of Molecular Probes, Inc.
| Catalog No. | Size |
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| CopyControl™ BAC Cloning Kit (BamH I) | |
| CCBAC1B | 1 Kit |
| For constructing the equivalent of one 10X human BAC library. | |
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| CopyControl™ BAC Cloning Kit (EcoR I) | |
| CCBAC1E | 1 Kit |
| For constructing the equivalent of one 10X human BAC library. | |
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| CopyControl™ BAC Cloning Kit (Hind III) | |
| CCBAC1H | 1 Kit |
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For constructing the equivalent of one 10X human BAC library. Contents: CopyControl™ pCC1BAC™ (BamH I, Hind III, or EcoR I) Cloning Ready Vector, Fast-Link™ DNA Ligase, Fast-Link™ 10X Ligation Buffer, ATP, BAC-Tracker™ Supercoiled DNA Ladder, EpiBlue™ Solution, EpiLyse™ Solution, Control Genomic DNA Insert, Control BAC Clone DNA (145 kb) |
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| CopyControl™ pCC1BAC™ (BamH I Cloning-Ready) Vector | |
| CBAC311B | 375 ng |
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Supplied at 25 ng/µl; vector is linearized at BamH I site and desphosphorylated. EPI300™ cells must be purchased separately. |
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| CopyControl™ pCC1BAC™ (EcoR I Cloning-Ready) Vector | |
| CBAC311E | 375 ng |
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Supplied at 25 ng/µl; vector is linearized at EcoR I site and desphosphorylated. EPI300™ cells must be purchased separately. |
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| CopyControl™ pCC1BAC™ (Hind III Cloning-Ready) Vector | |
| CBAC311H | 375 ng |
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Supplied at 25 ng/µl; vector is linearized at Hind III site and desphosphorylated. EPI300™ cells must be purchased separately. |
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| pCC1BAC/pIndigoBAC-5 Forward End-Sequencing Primer | |
| BFP0701 | 1 nmol |
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| pCC1BAC/pIndigoBAC-5 Reverse End-Sequencing Primer | |
| BRP0801 | 1 nmol |
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| CopyControl™ Induction Solution | |
| CCIS125 | 25 ml |
| 1,000X concentrated solution. Filter sterilized. | |
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| CopyControl™ BAC Autoinduction Solution | |
| AIS107B | 150 ml |
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