chiudiPCR cloning
GC Cloning (patents pending) is the first major advance in PCR cloning since the first description of TA-cloning (Mead, D., et. al. (1991) Biotechnology 9, 657.)
The GC Cloning technology is based on the discovery that these same enzymes add a single 3´-G to DNA molecules, either during PCR or as a separate G-tailing reaction to any blunt DNA.
GC Cloning and Amplification Kits (pGC™ Blue and pSMART® GC Vectors)
The pGC™ Blue Vector and The pSMART® GC vectors contains a single 3´-C overhang, which is compatible with the single 3´-G overhang on the inserts. The pGC Blue vector includes the lacZ sequence for blue/white screening, as well as dual opposed RNA polymerase promoters for in vitro transcription of the cloned insert.
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 Available Low and High copy number |
- Clone PCR products from proofreading or non-proofreading polymerases
- Stabilize otherwise unclonable inserts.
- No lost clones or deleted sequences.
- T7 and SP6 RNA polymerase promoters (for pGC™ Blue)
GC Cloning & Amplification Kits Content
4X GC Vector Premix (includes buffer, ATP, and either pGC™ Blue or pSMART® GC HK or pSMARTGC LK, CloneSmart® DNA Ligase, T4 Polynucleotide Kinase, 10X Primer Kinase Buffer (containing ATP), EconoTaq® DNA Polymerase, EconoTaq 10X Reaction Buffer (with Mg++), Control lacZ template plus PCR primers, CloneSmart Sequencing Primers; Recovery Medium, Control pUC DNA. A choice of E. cloni cells in SOLO packaging (one transformation per tube): Electrocompetent 10G ELITE 10G (> 2 x 1010 cfu/µg) or SUPREME (> 4 x 1010 cfu/µg) or Chemically Competent 10G (> 1 x 109 cfu/µg)
BigEasy® v2.0 Linear Cloning System (pJAZZ® Vectors)
The BigEasy Long PCR Cloning Kits contain everything needed to efficiently clone blunt or G-tailed PCR products up to 30 kb into the unbiased, high-fidelity pJAZZ linear cloning vector. There are two different versions of the vector (Figure 2) that are compatible with either proofreading (pJAZZ-OK Blunt Vector) or non-proofreading (pJAZZ-OK GC Vector) PCR polymerases.
The BigEasy Linear Cloning Kit is ideal for constructing bias-free, large-insert genomic libraries, or for cloning difficult DNA of any size up to 30 kb.
Note: pJAZZ-OC vector contains the Camr (Chloramphenicol resistance gene) instead of Kanr gene
- Clone any long or short PCR product.
- Clone PCR products from proofreading or non-proofreading polymerases.
- Stabilize otherwise unclonable inserts.
- Create libraries from A/T-rich or G/C-rich genomes.
- Clone gene clusters or operons
- Contain everything needed to efficiently clone blunt (pJAZZ-OK/OC Blunt Vector), sticky (pJAZZ-OK/OC NotI Vector) or G-tailed PCR products (pJAZZ-OK GC Vector) up to 30 kb into the unbiased, high-fidelity pJAZZ linear cloning vector
- A linear and not supercoiled vector allow unclonable sequences to be stabilized
- Transcription-free cloning technology, which further increases insert stability
- Inducible copy-number
BigEasy Long PCR Cloning Kit content
Pre-cut vector pJAZZ-OK (Blunt or C-tailed overhang), CloneSmart® DNA Ligase, CloneDirect 10X Ligation Buffer, PCR Control Template and Primers, Sequencing Primers, T4 Polynucleotide Kinase, 10X Primer Kinase Buffer, BigEasy-TSA Electrocompetent Cells, Positive Control Plasmid, Recovery Medium, Arabinose Induction Solution, and complete protocols.
BigEasy Linear Cloning Kit content
Dephosphorylated pJAZZ vector pre-cut at either a SmaI (blunt) or NotI site, CloneSmart DNA Ligase, CloneDirect™ 10X Ligation Buffer (includes ATP), Sequencing Primers, Positive Control Insert DNA, DNA Terminator® End Repair Enzyme, BigEasy-TSA Electrocompetent Cells in SOLO packaging (1 transformation per tube), Recovery Medium, Transformation Control DNA, and complete protocols. BigEasy-TSA Electrocompetent Cells are also available separately.
For more informations on cloning products, please click here
