CopyControl™ BAC Cloning Kit (BamH I)

Catalogue number

Pack size

CCBAC1B

1 kit

CopyControl™ BAC Cloning Kit (EcoR I)

Catalogue number

Pack size

CCBAC1E

1 kit

CopyControl™ BAC Cloning Kit (Hind III)

Catalogue number

Pack size

CCBAC1H

1 kit

CopyControl™ Induction Solution

Catalogue number

Pack size

CCIS125

25ml

CopyControl™ pCC1BAC™ (BamH I Cloning-Ready) Vector

Catalogue number

Pack size

CBAC311B

375ng

CopyControl™ pCC1BAC™ (EcoR I Cloning-Ready) Vector

Catalogue number

Pack size

CBAC311E

375ng

CopyControl™ pCC1BAC™ (Hind III Cloning-Ready) Vector

Catalogue number

Pack size

CBAC311H

375ng

pCC1BAC/pIndigoBAC-5 Forward End-Sequencing Primer

Catalogue number

Pack size

BFP0701

50mM

pCC1BAC/pIndigoBAC-5 Reverse End-Sequencing Primer

Catalogue number

Pack size

BRP0801

50mM

The CopyControl™ BAC Cloning Kits are the first commercially available kits for constructing BAC (Bacterial Artificial Chromosome) libraries. The kits have been designed to significantly reduce the time and labour normally required to construct and size a BAC library from any source of genomic DNA. The kits contain reagents sufficient for constructing and sizing the equivalent of one 10X human BAC library.

CopyControl™ BAC clones are initially selected and maintained as single copy clones to ensure their stability. Then, BAC clone copy number can be induced 10 - 20 fold for high yields of higher purity DNA for sequencing, fingerprinting and subcloning.

The kits feature the cloning ready CopyControl™ pCC1BAC™ vector which is derived from pBeloBAC11, pIndigoBAC, and Epicentre®'s pIndigoBAC-5 and contains both a single copy origin of replication and the inducible oriV multiple copy origin of replication. The CopyControl™ pCC1BAC™ (BamH I - Cloning Ready) and the CopyControl™ pCC1BAC™ (Hind III - Cloning Ready) vectors as provided in the kits are linearized at the unique BamH I or Hind III sites respectively, dephosphorylated and extensively purified and are ready for ligation. DNA ligation of the genomic DNA into the vectors can be completed in as little as 4 hours. Reagents and the BAC-Tracker™ Supercoiled DNA Ladder are provided in the kits to facilitate size screening of the BAC clones and for a rapid assessment of the quality of the BAC library in 3 hours. In addition, a control genomic DNA insert of 50kb and a control BAC clone of 145kb are provided to test ligation efficiency and transformation efficiency respectively.

TransforMax™ EC300™ Electrocompetent E. coli that are required for induction of the CopyControl™ BAC clones to multiple copy are supplied separately. The copy number of CopyControl™ BAC clones can be increased 10 - 20 fold when using TransforMax™ EC300™ cells.

Sequencing Primers for end-sequencing of CopyControl™ BAC clones are also available separately.

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.

Figure 2. Not I digestion of 2µl of 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.

Figure 3. The CopyControl™ BAC Cloning Kits significantly reduce the time and labor needed to construct a BAC library compared to standard methods.

Last modified: 2008-05-14 16:46:46

Benefits

• Kits include sufficient reagents for preparing and size screening the equivalent of a 10X human BAC library
• No need to prepare a BAC cloning vector - vectors are supplied linearized, dephosphorylated and highly purified
• Very low background. Greater than 95% white recombinant colonies
• DNA ligations in as little as 4 hours
• Screen the size of your BAC clone inserts in as little as 3 hours without the need to isolate DNA
• Induce a 10 - 20 fold increase in copy number of CopyControl BAC clone for high yields of higher purity DNA
• End-sequence and fingerprint your CopyControl™ BAC clones from a single miniprep

Last modified: 2008-05-14 16:46:46

Protocols for: CopyControl™ BAC Cloning Kits (BamH, EcoR I, Hind III) & CopyControl™ pCC1BAC™ Cloning-Ready Vectors (BamH, EcoR I, Hind III)

CopyControl™ BAC Cloning Kits Protocol

(catalogue number CCBAC1B / CCBAC1E / CCBAC1H)

CopyControl™ pCC1BAC™ Cloning-Ready Vectors Protocol

(catalogue number CBAC311B / CBAC311E / CBAC311H)

Please note: all protocols off site are the responsibility of the products supplier

Last modified: 2008-05-15 10:43:29